FIELD OF THE INVENTION

The present invention provides polynucleotides encoding polypeptides associated with the development and progression of rheumatoid arthritis and homologs thereof. Also provided are vectors, host cells, antibodies, and recombinant and syntheticmethods for producing said polypeptides. The invention further relates to diagnostic and therapeutic methods for utilizing these polypeptides in the diagnosis, treatment, and/or prevention of rheumatoid arthritis and related disease states. Theinvention further relates to screening methods for identifying agonists and antagonists of the polynucleotides and polypeptides of the present invention.

BACKGROUND OF RELATED TECHNOLOGY

Rheumatoid arthritis (RA) is a chronic inflammatory disease characterized by progressive joint destruction. Initial destruction of cartilage and bone is associated with the formation of a pannus, consisting of a hypertrophic synovial membranecontaining hyperplastic synoviocytes and an infiltrate of inflammatory cells including T cells, B cells, CD68+ macrophages, mast cells, and endothelial cells. The causes of RA are not well understood. Genetic studies have linked expression of specificmajor histocompatibility complex class II antigens to the development of RA, suggesting the involvement of antigen-specific mechanisms in disease progression (Zanelli et al., Hum. Immunol. 61:1254-1261 (2000)).

CD4+ T cells are thought to play a key role in initiation and progression of disease. Although many putative self antigens have been proposed, none have been definitively associated with the initiation of disease. Antigen-activated T cellsstimulate monocytes, macrophages, and synovial fibroblasts to secrete pro-inflammatory cytokines including interleukin-1 (IL-1), interleukin-6 (IL-6), and TNF-.alpha.. These cytokines stimulate synovial fibroblasts, osteoclasts, and chondrocytes torelease matrix metalloproteinases (MMPs) that destroy surrounding tissue. Activated CD4+ T cells stimulate osteoclastogenesis that can also contribute to joint damage. The activated T cells also stimulate B cells present in the synovium via the CD40pathway to differentiate into antibody secreting cells producing rheumatoid factor, which may also contribute to disease pathology.

Many of the cytokines found in rheumatoid synovium have been directly linked to disease pathology. For example, TNF-.alpha. promotes inflammation by inducing secretion of other inflammatory cytokines including IL-1, IL-6, IL-8, GM-CSF, as wellas by upregulating adhesion molecule expression on endothelial cells and synovial fibroblasts. These two events promote increased migration of lymphocytes including neutrophils, monocytes, and T cells into the synovium. Neutrophils release elastase andproteases that degrade proteoglycan and contribute to joint destruction. Therapies targeting TNF-.alpha. include the use of soluble TNF-.alpha. receptor (Etanercept) and neutralizing antibodies specific for TNF-.alpha. (Infliximab), and result in asignificant decrease in the number of swollen joints, as well as the numbers of T cells and plasma cells in the synovium of RA patients. Such therapies also result in a decrease in the expression of VCAM-1 and IL-1 in the synovium of treated patients(Bathon, et al., New Engl. J. Med. 343:1586-1593 (2000); Lipsky, et al., New Engl. J. Med. 343:1594-1602 (2000); Richard-Miceli, et al., Biodrugs 15:251-259 (2001)).

IL-1 has also been closely linked to the pathophysiology of RA. IL-1 induces synovial cell proliferation and activates MMP and prostaglandin production in vitro (Mizel et al., Proc. Natl. Acad. Sci. USA 78:2474-2477 (1981)). In severalmouse models of arthritis, IL-1 is believed to play a dominant role in cartilage destruction, whereas TNF-.alpha. is primarily proinflammatory (Joosten et al., J. Immunol. 163:5049-5055 (1999)). Transgenic mice constitutively expressing humanIL-1.alpha. in various organs develop a severe polyarthritic phenotype with a predominance of neutrophils and macrophages in the diseased joints (Niki et al., J. Clin. Invest. 107:1127-1135 (2001)). Synovitis developed within two weeks of birth,followed by pannus formation and cartilage destruction within 8 weeks after birth. Treatment of RA patients with a natural inhibitor of IL-1, recombinant human IL-1 receptor antagonist (IL-1Ra), significantly reduced clinical symptoms and the rate ofprogressive joint damage (Jiang et al., Arthritis Rheum. 43:1001-1009 (2000); Bresnihan et al., Biodrugs 15:87-97 (2001)).

A number of studies have sought to identify genes whose expression is associated with the development of RA. cDNA microarrays have been used to compare expression profiles between tissue samples derived from RA and inflammatory bowel diseasepatients. Such studies have found that prominently upregulated genes in RA samples include: IL-6; the MMPs stromelysin-1, collagenase-1, gelatinase A, and human matrix metalloelastase; tissue inhibitors of metalloproteinases, including TIMP-1 andTIMP-3; the adhesion molecule VCAM-1; and chemokines including MCP-1, MIF, and RANTES (Heller et al. Proc. Natl. Acad. Sci. USA 94: 2150-2155 (1997)). Further, a cDNA library has been generated from monocytes obtained from a RA patient with activedisease (Stuhlmuller et al., Arthritis Rheum. 43:775-790 (2000)). Genes found to be upregulated in these cells include IL-1.alpha., IL-1.beta., IL-6, TNF-.alpha., growth-related oncogene .alpha., macrophage inflammatory protein 2, ferritin,.alpha.1-antitrypsin, lysozyme, transaldolase, Epstein-Barr virus-encoded RNA 1-associated protein, thrombospondin 1, angiotensin receptor II C-terminal homologue, and RNA polymerase II elongation factor.

In one study, a cDNA library was generated by subtracting cDNA derived from noninflammatory osteoarthritis (OA) synoviocytes from cDNA derived from cultured RA fibroblastoid synoviocytes (Seki et al., Arthritis Rheum. 41:1356-1364 (1998)). Genes found to be constitutively overexpressed in the rheumatoid synoviocyte line include: chemokine stromal cell-derived factor 1.alpha.; adhesion molecule VCAM-1; interferon-inducible 56-kD protein; 2'-5'-oligoadenylate synthetase; Mac-2 bindingprotein; extracellular matrix components biglycan, lumican, and IGFBP5; and semaphorin VI.

Studies have also been conducted using suppression subtractive hybridization to identify genes that are highly expressed in RA synovium relative to OA synovium (Justen et al., Mol. Cell Biol. Res. Comm. 3:165-172 (2000)). Genes found to bespecifically upregulated in RA synovium include: cytoskeletal .gamma.-actin; the extracellular matrix components fibronectin and collagen III.alpha.1; superficial zone protein; elongation factor .alpha.1; granulin precursor; interferon-.gamma. induciblelysosomal thiol reductase; the protease cathepsin B; phospholipase A2 group IIA; and annexin II.

Accordingly, there is a continuing need to identify genes whose expression is associated with the development and progression of RA. The identification of such genes permits the development of clones expressing such genes, thereby permitting theidentification of compounds capable of modulating the activity of such genes and/or their expression products. Such compounds may have therapeutic utility in the diagnosis and/or treatment of RA and related disease states. The present invention isdirected to meeting these and other needs.

BRIEF DESCRIPTION OF THE DRAWINGS

FIGS. 1, 4, 7, 10, 13, 16, 19, 22, 25, 28, 31, 34, 37, 40, 43, 46, 49, 52, 55, 58, 61, 64, 67, 69, 71, 74, 76, 79, 82 and 85 show microarray data for genes of the present invention shown to be upregulated and downregulated in rheumatoid arthritissynovial fluid.

FIGS. 2, 5, 8, 11, 14, 17, 20, 23, 26, 29, 32, 35, 38, 41, 44, 47, 50, 53, 56, 59, 62, 65, 68, 70, 72, 75, 77, 80, 83 and 86 show polynucleotide sequences for genes of the present invention shown to be upregulated and downregulated in rheumatoidarthritis synovial fluid.

FIGS. 3, 6, 9, 12, 15, 18, 21, 24, 27, 30, 33, 36, 39, 42, 45, 48, 51, 54, 57, 60, 63, 66, 73, 78, 81, 84 and 87 show amino acid sequences for the expression product of genes of the present invention shown to be upregulated and downregulated inrheumatoid arthritis synovial fluid.

FIG. 88 shows the regulation of GBP-1 and GBP-5 expression by NFkB.

FIG. 89 shows the time course of GBP-1 and GBP-5 expression.

FIG. 90 shows GBP-1 and GBP-5 expression in mouse embryonic fibroblast lines derived from NFkB and IkB.alpha. germline knockouts.

FIG. 91 shows tissue expression patterns of GBP-1 and GBP-5.

FIG. 92 shows expression of GBP-1 and GBP-5 in resting and stimulated THP-1 monocytes.

FIG. 93 shows expression of GBP-1 and GBP-5 in resting and stimulated human microvascular endothelial cells.

FIG. 94 shows expression of GBP-1 and GBP-5 in resting and stimulated fibroblasts derived from rheumatoid arthritis synovium.

FIG. 95 shows expression of GBP-1 and GBP-5 in resting and stimulated peripheral blood T cells.

SUMMARY OF THE INVENTION

In one aspect, the present invention is directed to an assay for identifying a compound that modulates the activity of a gene associated with rheumatoid arthritis, including the steps of: (1) providing a cell expressing a gene associated withrheumatoid arthritis, wherein the nucleic acid sequence of the gene associated with rheumatoid arthritis is at least 95% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NO:37, SEQ ID NO:39, SEQ ID NO:41, SEQ ID NO:43,SEQ ID NO:45, SEQ ID NO:47, SEQ ID NO:49, SEQ ID NO:51, SEQ ID NO:53, SEQ ID NO:55, SEQ ID NO:57, SEQ ID NO:59, SEQ ID NO:61, SEQ ID NO:63, SEQ ID NO:65, SEQ ID NO:67, SEQ ID NO:69, SEQ ID NO:70, SEQ ID NO:71, SEQ ID NO:73, and SEQ ID NO:74; (2)contacting the cell expressing the gene associated with rheumatoid arthritis with a test compound; and (3) determining whether the test compound modulates the activity of the gene associated with rheumatoid arthritis. The assay may be cell-based assayor may be a cell-free assay, such as a ligand-binding assay. The test compound desirably modulates the activity of the gene associated with rheumatoid arthritis, may be an antagonist or an agonist of the gene associated with rheumatoid arthritis, andmay bind to the gene associated with rheumatoid arthritis. The assay is desirably useful for identifying compounds which are useful for the treatment of rheumatoid arthritis.

In another aspect, the present invention is directed to an assay for identifying a compound that modulates the activity of a protein associated with rheumatoid arthritis, including the steps of: (1) providing a cell expressing a gene associatedwith rheumatoid arthritis, wherein the gene encodes a polypeptide having an amino acid sequence which is at least 95% identical to an amino acid sequence selected from the group consisting of SEQ ID NO:38, SEQ ID NO:40, SEQ ID NO:42, SEQ ID NO:44, SEQ IDNO:46, SEQ ID NO:48, SEQ ID NO:50, SEQ ID NO:52, SEQ ID NO:54, SEQ ID NO:56, SEQ ID NO:58, SEQ ID NO:60, SEQ ID NO:62, SEQ ID NO:64, SEQ ID NO:66, SEQ ID NO:68, SEQ ID NO:72, and SEQ ID NO:75; (2) contacting the cell expressing the gene associated withrheumatoid arthritis with a test compound; and (3) determining whether the test compound modulates the activity of the protein associated with rheumatoid arthritis. The test compound desirably modulates the activity of the protein associated withrheumatoid arthritis, may be an antagonist or an agonist of the protein associated with rheumatoid arthritis, and may bind to the protein associated with rheumatoid arthritis. The assay is desirably useful for identifying compounds which are useful forthe treatment of rheumatoid arthritis.

In another aspect, the present invention is directed to a method for the treatment of rheumatoid arthritis, including the steps of: (1) identifying a patient suffering from rheumatoid arthritis; and (2) administering to the patient atherapeutically effective amount of a modulator of a gene associated with rheumatoid arthritis, wherein the gene associated with rheumatoid arthritis has a nucleic acid sequence which is at least 95% identical to a nucleic acid sequence selected from thegroup consisting of SEQ ID NO:37, SEQ ID NO:39, SEQ ID NO:41, SEQ ID NO:43, SEQ ID NO:45, SEQ ID NO:47, SEQ ID NO:49, SEQ ID NO:51, SEQ ID NO:53, SEQ ID NO:55, SEQ ID NO:57, SEQ ID NO:59, SEQ ID NO:61, SEQ ID NO:63, SEQ ID NO:65, SEQ ID NO:67, SEQ IDNO:69, SEQ ID NO:70, SEQ ID NO:71, SEQ ID NO:73, and SEQ ID NO:74. The patient is desirably identified as suffering from rheumatoid arthritis by measuring the expression level of the gene associated with rheumatoid arthritis in the patient. Themodulator is desirably an antagonist of a gene associated with rheumatoid arthritis.

In another aspect, the present invention is directed to a method for the treatment of rheumatoid arthritis, including the steps of: (1) identifying a patient suffering from rheumatoid arthritis; and (2) administering to the patient suffering fromrheumatoid arthritis a therapeutically effective amount of a modulator of a polypeptide associated with rheumatoid arthritis, wherein the polypeptide associated with rheumatoid arthritis has an amino acid sequence which is at least 95% identical to anamino acid sequence selected from the group consisting of SEQ ID NO:38, SEQ ID NO:40, SEQ ID NO:42, SEQ ID NO:44, SEQ ID NO:46, SEQ ID NO:48, SEQ ID NO:50, SEQ ID NO:52, SEQ ID NO:54, SEQ ID NO:56, SEQ ID NO:58, SEQ ID NO:60, SEQ ID NO:62, SEQ ID NO:64,SEQ ID NO:66, SEQ ID NO:68, SEQ ID NO:72, and SEQ ID NO:75. The patient is desirably identified as suffering from rheumatoid arthritis by measuring the expression level of the polypeptide associated with rheumatoid arthritis. The modulator is desirablyan antagonist of a polypeptide associated with rheumatoid arthritis.

In another aspect, the present invention is directed to a method for the prevention of rheumatoid arthritis, including the steps of: (1) identifying a patient at risk for rheumatoid arthritis; and (2) administering to the patient at risk forrheumatoid arthritis a therapeutically effective amount of a modulator of a gene associated with rheumatoid arthritis, wherein the gene associated with rheumatoid arthritis has a nucleic acid sequence which is at least 95% identical to a nucleic acidsequence selected from the group consisting of SEQ ID NO:37, SEQ ID NO:39, SEQ ID NO:41, SEQ ID NO:43, SEQ ID NO:45, SEQ ID NO:47, SEQ ID NO:49, SEQ ID NO:51, SEQ ID NO:53, SEQ ID NO:55, SEQ ID NO:57, SEQ ID NO:59, SEQ ID NO:61, SEQ ID NO:63, SEQ IDNO:65, SEQ ID NO:67, SEQ ID NO:69, SEQ ID NO:70, SEQ ID NO:71, SEQ ID NO:73, and SEQ ID NO:74. The patient is desirably identified as being at risk for rheumatoid arthritis by measuring the expression level of the gene associated with rheumatoidarthritis in the patient.

In another aspect, the present invention is directed to a method for the prevention of rheumatoid arthritis, including the steps of: (1) identifying a patient at risk for rheumatoid arthritis; and (2) administering to the patient at risk forrheumatoid arthritis a therapeutically effective amount of a modulator of a polypeptide associated with rheumatoid arthritis, wherein the polypeptide associated with rheumatoid arthritis has an amino acid sequence which is at least 95% identical to anamino acid sequence selected from the group consisting of SEQ ID NO:38, SEQ ID NO:40, SEQ ID NO:42, SEQ ID NO:44, SEQ ID NO:46, SEQ ID NO:48, SEQ ID NO:50, SEQ ID NO:52, SEQ ID NO:54, SEQ ID NO:56, SEQ ID NO:58, SEQ ID NO:60, SEQ ID NO:62, SEQ ID NO:64,SEQ ID NO:66, SEQ ID NO:68, SEQ ID NO:72, and SEQ ID NO:75. The patient is desirably identified as being at risk for rheumatoid arthritis by measuring the expression level of the polypeptide associated with rheumatoid arthritis in the patient.

In another aspect, the present invention is directed to a compound useful for the treatment of rheumatoid arthritis, wherein the compound is identified by: (1) providing a cell expressing a gene associated with rheumatoid arthritis, wherein thegene associated with rheumatoid arthritis has a nucleic acid sequence which is at least 95% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NO:37, SEQ ID NO:39, SEQ ID NO:41, SEQ ID NO:43, SEQ ID NO:45, SEQ ID NO:47, SEQID NO:49, SEQ ID NO:51, SEQ ID NO:53, SEQ ID NO:55, SEQ ID NO:57, SEQ ID NO:59, SEQ ID NO:61, SEQ ID NO:63, SEQ ID NO:65, SEQ ID NO:67, SEQ ID NO:69, SEQ ID NO:70, SEQ ID NO:71, SEQ ID NO:73, and SEQ ID NO:74; (2) contacting the cell expressing the geneassociated with rheumatoid arthritis with the compound; and (3) determining whether the compound modulates the activity of the gene associated with rheumatoid arthritis.

In another aspect, the present invention is directed to a compound useful for the treatment of rheumatoid arthritis, wherein the compound is identified by: (1) providing a cell expressing a polypeptide associated with rheumatoid arthritis,wherein the polypeptide associated with rheumatoid arthritis has an amino acid sequence which is at least 95% identical to an amino acid sequence selected from the group consisting of SEQ ID NO:38, SEQ ID NO:40, SEQ ID NO:42, SEQ ID NO:44, SEQ ID NO:46,SEQ ID NO:48, SEQ ID NO:50, SEQ ID NO:52, SEQ ID NO:54, SEQ ID NO:56, SEQ ID NO:58, SEQ ID NO:60, SEQ ID NO:62, SEQ ID NO:64, SEQ ID NO:66, SEQ ID NO:68, SEQ ID NO:72, and SEQ ID NO:75; (2) contacting the cell expressing the polypeptide associated withrheumatoid arthritis with the compound; and (3) determining whether the compound modulates the activity of the polypeptide associated with rheumatoid arthritis.

DETAILED DESCRIPTION OF THE INVENTION

The present invention is directed to the identification of genes associated with Rheumatoid Arthritis (RA). Such genes and their polypeptide expression products are hereinafter referred to as "RA-associated genes and polypeptides". In thepresent invention, RA-associated genes and polypeptides have been identified by probing Affymetrix chips (describe) with mRNA derived from the synovia of RA patients, as set forth in Section A of "Materials and Methods", hereinbelow. Gene expressionpatterns were compared to those obtained using mRNA derived from synovia of control joint trauma patients. Several genes were identified as having significantly increased expression in the RA synovium relative to the controls, as further describedhereinbelow. Several genes have also been identified as having significantly decreased expression in the RA synovium relative to the controls, as further described hereinbelow.

The present invention provides synthetic methods for producing RA-associated genes and polypeptides. Also provided are diagnostic methods for detecting diseases, disorders, and/or conditions related to RA-associated genes and polypeptides, andtherapeutic methods for treating such diseases, disorders, and/or conditions. The invention further relates to screening methods for identifying binding partners of RA-associated genes and polypeptides.

Examples of functional assays useful in the present invention include LPS-induced TNF.alpha. and TNF.alpha.-induced IL-1.beta. secretion by THP-1 monocytes, anti-CD3/anti-CD28-induced IL-2 secretion by Jurkat T cells, TNF.alpha.-inducedIL-1.beta. secretion by synovial fibroblasts, TNF.alpha.-induced E-selectin expression by endothelial cells, and anti-CD40-induced homotypic aggregation of Raji B cells.

One of skill in the art will recognize that RA-associated genes and polypeptides of the present invention are desirably murine or human, but may be from any suitable organism. The genomic and protein sequences of RA-associated genes andpolypeptides from these organisms are readily accessed via Genbank or The National Center for Biotechnology Information.

Further, derivatives and homologues of RA-associated genes and polypeptides may be used in the present invention. For example, nucleic acid sequences of RA-associated genes of the present invention may be altered by substitutions, additions, ordeletions that provide for functionally equivalent-conservative variants of such genes. Further, one or more amino acid residues within the amino acid sequence of RA-associated polypeptides can be substituted by another amino acid of similar properties,such as, for example, positively charged amino acids (arginine, lysine, and histidine); negatively charged amino acids (aspartate and glutamate); polar neutral amino acids; and non-polar amino acids.

Other conservative amino acid substitutions can be taken from the Table 1, below.

TABLE-US-00001 TABLE 1 Conservative Amino Acid Replacements For Amino Acid Code Replace with any of: Alanine A D-Ala, Gly, beta-Ala, L-Cys, D-Cys Arginine R D-Arg, Lys, D-Lys, homo-Arg, D-homo-Arg, Met, Ile, D-Met, D-Ile, Orn, D-Orn Asparagine ND-Asn, Asp, D-Asp, Glu, D-Glu, Gln, D-Gln Aspartic Acid D D-Asp, D-Asn, Asn, Glu, D-Glu, Gln, D-Gln Cysteine C D-Cys, S-Me-Cys, Met, D-Met, Thr, D-Thr Glutamine Q D-Gln, Asn, D-Asn, Glu, D-Glu, Asp, D-Asp Glutamic Acid E D-Glu, D-Asp, Asp, Asn, D-Asn,Gln, D-Gln Glycine G Ala, D-Ala, Pro, D-Pro, .beta.-Ala, Acp Isoleucine I D-Ile, Val, D-Val, Leu, D-Leu, Met, D-Met Leucine L D-Leu, Val, D-Val, Met, D-Met Lysine K D-Lys, Arg, D-Arg, homo-Arg, D-homo-Arg, Met, D-Met, Ile, D-Ile, Orn, D-Orn Methionine MD-Met, S-Me-Cys, Ile, D-Ile, Leu, D-Leu, Val, D-Val Phenylalanine F D-Phe, Tyr, D-Thr, L-Dopa, His, D-His, Trp, D-Trp, Trans-3,4, or 5-phenylproline, cis-3,4, or 5-phenylproline Proline P D-Pro, L-1-thioazolidine-4-carboxylic acid, D- orL-1-oxazolidine-4-carboxylic acid Serine S D-Ser, Thr, D-Thr, allo-Thr, Met, D-Met, Met(O), D-Met(O), L-Cys, D-Cys Threonine T D-Thr, Ser, D-Ser, allo-Thr, Met, D-Met, Met(O), D-Met(O), Val, D-Val Tyrosine Y D-Tyr, Phe, D-Phe, L-Dopa, His, D-His ValineV D-Val, Leu, D-Leu, Ile, D-Ile, Met, D-Met

Other analogs within the present invention are those with modifications which increase protein stability; such analogs may contain, for example, one or more non-peptide bonds (which replace the peptide bonds) in the protein sequence. Alsoincluded are analogs that include residues other than naturally occurring L-amino acids, e.g., D-amino acids or non-naturally occurring or synthetic amino acids, e.g., .beta. or .gamma. amino acids.

RA-associated polypeptides of the present invention may be modified by, for example, phosphorylation, sulfation, acylation, or other protein modifications. They may also be modified with a label capable of providing a detectable signal, eitherdirectly or indirectly, including, but not limited to, radioisotopes and fluorescent compounds.

It will be apparent to one of skill in the art that conventional screening assays may be used in methods of the present invention for the identification of modulators of RA-associated genes and polypeptides.

In the present invention, techniques for screening large gene libraries may include cloning the gene library into replicable expression vectors, transforming appropriate cells with the resulting library of vectors, and expressing the genes underconditions for detection of a desired activity. Techniques known in the art are amenable to high throughput analysis for screening large numbers of sequences created, e.g., by random mutagenesis techniques. High throughput assays can be followed bysecondary screens in order to identify further biological activities which will, e.g., allow one skilled in the art to differentiate agonists from antagonists. The type of a secondary screen used will depend on the desired activity that needs to betested.

Drug screening assays are also provided in the present invention. By producing purified and recombinant forms of RA-associated genes and polypeptides of the present invention, or fragments thereof, one skilled in the art can use these to screenfor drugs which are either agonists or antagonists of the normal cellular function or their role in cellular signaling. In one aspect, the assay evaluates the ability of a compound to modulate binding between RA-associated genes and polypeptides of thepresent invention and a naturally occurring ligand. The term "modulating" encompasses enhancement, diminishment, activation or inactivation of activity of RA-associated genes and polypeptides. Assays useful for identifying ligands to RA-associatedgenes and polypeptides of the present invention are encompassed herein. Such ligands include peptides, proteins, small molecules, and antibodies, which are capable of binding to RA-associated genes and polypeptides of the present invention andmodulating their activity. A variety of assay formats may be used in the present invention and are known by those skilled in the art.

In many drug screening programs which test libraries of compounds and natural extracts, high throughput assays are desirable in order to maximize the number of compounds surveyed in a given period of time. Assays which are performed in cell-freesystems, such as may be derived with purified or semi-purified proteins, are often preferred as primary screens in that they can be generated to permit rapid development and relatively easy detection of an alteration in a molecular target which ismediated by a test compound.

Compounds identified using assays, as discussed hereinabove, may be antagonists or agonists of RA-associated genes and polypeptides. These compounds are useful in modulating the activity of RA-associated genes and polypeptides and in treatingdisorders associated with RA-associated genes and polypeptides.

"Disorders associated with RA-associated genes and polypeptides" refers to any disorder or disease state in which RA-associated genes and polypeptides play a regulatory role in the metabolic pathway of that disorder or disease. As used herein,the term "treating" refers to the alleviation of symptoms of a particular disorder in a patient, the improvement of an ascertainable measurement associated with a particular disorder, or the prevention of a particular immune, inflammatory or cellularresponse.

A compound which acts as a modulator of RA-associated genes and polypeptides may be administered for therapeutic use as a raw chemical or may be the active ingredient in a pharmaceutical formulation. Such formulations of the present inventionmay contain other therapeutic agents as described below, and may be formulated, for example, by employing conventional solid or liquid vehicles or diluents, as well as pharmaceutical additives of a type appropriate to the mode of desired administration(for example, excipients, binders, preservatives, stabilizers, flavors, etc.) according to techniques such as those well known in the art of pharmaceutical formulation.

Compounds of the present invention may be administered by any suitable means, for example, orally, such as in the form of tablets, capsules, granules or powders; sublingually; buccally; parenterally, such as by subcutaneous, intravenous,intramuscular, or intrasternal injection or infusion techniques (e.g., as sterile injectable aqueous or non-aqueous solutions or suspensions); nasally such as by inhalation spray; topically, such as in the form of a cream or ointment; or rectally such asin the form of suppositories; in dosage unit formulations containing non-toxic, pharmaceutically acceptable vehicles or diluents.

Such compounds may, for example, be administered in a form suitable for immediate release or extended release. Immediate release or extended release may be achieved by the use of suitable pharmaceutical compositions comprising compounds of thepresent invention, or, particularly in the case of extended release, by the use of devices such as subcutaneous implants or osmotic pumps. Compounds of the present invention may also be administered liposomally.

Exemplary compositions for oral administration include suspensions which may contain, for example, microcrystalline cellulose for imparting bulk, alginic acid or sodium alginate as a suspending agent, methylcellulose as a viscosity enhancer, andsweeteners or flavoring agents such as those known in the art; and immediate release tablets which may contain, for example, microcrystalline cellulose, dicalcium phosphate, starch, magnesium stearate and/or lactose and/or other excipients, binders,extenders, disintegrants, diluents and lubricants such as those known in the art.

Compounds of the present invention may also be delivered through the oral cavity by sublingual and/or buccal administration. Molded tablets, compressed tablets or freeze-dried tablets are exemplary forms which may be used. Exemplarycompositions include those formulating the compound(s) of the present invention with fast dissolving diluents such as mannitol, lactose, sucrose and/or cyclodextrins.

Also included in such formulations may be high molecular weight excipients such as celluloses (avicel) or polyethylene glycols (PEG). Such formulations may also include an excipient to aid mucosal adhesion such as hydroxy propyl cellulose (HPC),hydroxy propyl methyl cellulose (HPMC), sodium carboxy methyl cellulose (SCMC), maleic anhydride copolymer (e.g., Gantrez), and agents to control release such as polyacrylic copolymer (e.g., Carbopol 934). Lubricants, glidants, flavors, coloring agentsand stabilizers may also be added for ease of fabrication and use.

Exemplary compositions for nasal aerosol or inhalation administration include solutions in saline which may contain, for example, benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, and/or othersolubilizing or dispersing agents such as those known in the art.

Exemplary compositions for parenteral administration include injectable solutions or suspensions which may contain, for example, suitable non-toxic, parenterally acceptable diluents or solvents, such as mannitol, 1,3-butanediol, water, Ringer'ssolution, an isotonic sodium chloride solution, or other suitable dispersing or wetting and suspending agents, including synthetic mono- or diglycerides, and fatty acids, including oleic acid.

Exemplary compositions for rectal administration include suppositories which may contain, for example, a suitable non-irritating excipient, such as cocoa butter, synthetic glyceride esters or polyethylene glycols, which are solid at ordinarytemperatures, but liquefy and/or dissolve in the rectal cavity to release the drug.

Exemplary compositions for topical administration include a topical carrier such as Plastibase (mineral oil gelled with polyethylene).

The effective amount of a compound of the present invention may be determined by one of ordinary skill in the art, and includes exemplary dosage amounts for an adult human of from about 0.1 to 100 mg/kg of body weight of active compound per day,which may be administered in a single dose or in the form of individual divided doses, such as from 1 to 4 times per day. It will be understood that the specific dose level and frequency of dosage for any particular subject may be varied and will dependupon a variety of factors including the activity of the specific compound employed, the metabolic stability and length of action of that compound, the species, age, body weight, general health, sex and diet of the subject, the mode and time ofadministration, rate of excretion, drug combination, and severity of the particular condition. Preferred subjects for treatment include animals, most preferably mammalian species such as humans, and domestic animals such as dogs, cats and the like,subject to disorders associated with RA-associated genes and polypeptides.

The compounds of the present invention may be employed alone or in combination with each other and/or other suitable therapeutic agents useful in the treatment of disorders associated with RA-associated genes and polypeptides.

In another aspect, the present invention relates to the use of an isolated nucleic acid in "antisense" therapy. As used herein, "antisense" therapy refers to administration or in situ generation of oligonucleotides or their derivatives whichspecifically hybridize under cellular conditions with the cellular mRNA and/or genomic DNA of RA-associated genes so as to inhibit expression of the proteins encoded by such genes, e.g., by inhibiting transcription and/or translation. In general,"antisense" therapy refers to the range of techniques generally employed in the art, and includes any therapy which relies on specific binding to oligonucleotide sequences.

Gene constructs useful in antisense therapy may be administered may be administered in any biologically effective carrier, e.g., any formulation or composition capable of effectively delivering a nucleic acid sequence to cells in vivo. Approaches include insertion of the subject gene in viral vectors including recombinant retroviruses, adenoviruses, adeno-associated viruses, and herpes simplex virus-1, or recombinant bacterial or eukaryotic plasmids. Viral vectors transfect cellsdirectly; an advantage of infection of cells with a viral vector is that a large proportion of the targeted cells can receive the nucleic acid. Several viral delivery systems are known in the art and can be utilized by one practicing the presentinvention.

In addition to viral transfer methods, non-viral methods may also be employed. Most non-viral methods of gene transfer rely on normal mechanisms used by mammalian cells for the uptake and intracellular transport of macromolecules. Exemplarygene delivery systems of this type include liposomal derived systems, poly-lysine conjugates, and artificial viral envelopes. Nucleic acid sequences may also be introduced to cell(s) by direct injection of the gene construct or by electroporation.

In clinical settings, the gene delivery systems can be introduced into a patient by any of a number of methods, each of which is known in the art. For instance, a pharmaceutical preparation of the gene delivery system can be introducedsystemically, e.g., by intravenous injection, and specific transduction of the protein in the target cells occurs predominantly from specificity of transfection provided by the gene delivery vehicle, cell-type or tissue-type expression due to thetranscriptional regulatory sequences controlling expression of the receptor gene, or a combination thereof.

The pharmaceutical preparation of the gene therapy construct can consist essentially of the gene delivery system in an acceptable diluent, or can comprise a slow release matrix in which the gene delivery vehicle is embedded. Alternatively, wherethe complete gene delivery system can be produced intact from recombinant cells, e.g., retroviral vectors, the pharmaceutical preparation can comprise one or more cells which produce the gene delivery system.

The following sections sets forth the materials and methods utilized in the present invention.

Materials and Methods

A. Microarray Experimentation

1. RNA Isolation

Human knee biopsy samples were homogenized in 3 ml TRIZOL.RTM. Reagent (Life Technologies, Rockville, Md.) and frozen in liquid nitrogen. The samples were thawed, one-third (1 ml) of the sample was removed and mixed with 1 ml TRIZOL.RTM.. Themixture was then homogenized and snap frozen in liquid nitrogen. Following a thaw, the samples were spun at 14,000 rpm for 10 minutes at 4.degree. C. The supernatants were transferred to new microfuge tubes, extracted with chloroform, and precipitatedwith isopropanol overnight at -20.degree. C. The RNA was pelleted by centrifugation at 14,000 rpm for 30 minutes. The supernatant was aspirated, and the samples washed two times with 75% ethanol. Following the last spin, the pellets were air-dried,and resuspended in 20 ul of ultra-pure RNase-free water. The RNA samples were further purified using Qiagen RNease mini columns (Qiagen Inc., Valencia Calif.), according to manufacturer's instructions. The RNA was eluted with 50 ul of RNase-free water.

2. Probe Preparation

The RNA was treated in a total reaction volume of 100 ul with RNase Inhibitor (Invitrogen Corp., Carlsbad, Calif.), DNase I (Ambion, Houston, Tex.) for 30 minutes at 37.degree. C. The treated RNA was purified using Qiagen RNease mini columns,according to the manufacturer's instructions.

For the first strand cDNA synthesis, the RNA was incubated with T7-(dT)24 primer, having the sequence: 5'GGCCAGTGAATTGTAATACGACTCACTATAGGGAGGCGGTTTTTTTT TTTTTTTTTTTTTTTT3'(SEQ ID NO:1) for 10 minutes at 70.degree. C., followed by one minute onice. First strand buffer, DTT, dNTP and RNase were added, and the samples incubated for 2 minutes at 45.degree. C. Superscript II reverse transcriptase (Invitrogen Corp, Carlsbad, Calif.) was added, and the samples incubated for an additional 60minutes at 45.degree. C.

For the second strand synthesis, the first strand cDNA was incubated with second strand buffer, dNTPs, E. coli ligase, E. coli RNase H, E. coli Polymerase I in a total volume of 150 ul for two hours at 16.degree. C. T4 polymerase was added, andthe incubation continued for an additional 5 minutes. Following this incubation, EDTA was added, and the samples placed on ice. The cDNA samples were extracted with phenol:chloroform:isoamyl alcohol and precipitated by addition of 0.5 volumes of 7.5 Mammonium acetate and 2.5 volumes of 100% ethanol. The samples were pelleted by a 30 minute room temperature spin at 12,000.times.g. The pelleted samples were washed with 0.5 ml 80% ethanol, spun for 10 minutes at 12,000.times.g, and air dried. Thesamples were resuspended in 12 ul RNase free water.

The cDNA was labeled using the Enzo Bio Array High Yield RNA transcript labeling kit (Enzo Therapeutics, Farmingdale, N.Y.). The cDNA was incubated with HY reaction buffer, biotin labeled NTP, DTT, RNase mix, and T7 DNA polymerase for six hoursat 37.degree. C. Unincorporated nucleotides were removed using Qiagen RNeasy columns according to manufacturer's instructions. The cRNA was fragmented by addition of fragmentation buffer, and incubated for 35 minutes at 95.degree. C. The fragmentedcRNA (0.05 mg/ml) was added to a hybridization solution master mix that included 0.1 mg/ml herring sperm DNA, 5 nM oligo B2, 1.times. standard curve pool, 0.5 mg/ml acetylated BSA, 1.times.MES hybridization buffer.

The Affymetrix human U95v2 A, B, and C GeneChips.RTM. were probed with the hybridization master mix. The hybridization, washing, and Phycoerythrin streptavidin staining were performed using the Affymetrix hybridization oven and fluidicsworkstation according to manufacturer's instructions. Stained chips were scanned on the Affymetrix GeneChip scanner, and data was analyzed using the Affymetrix GeneChip software to determine the specifically hybridizing signal for each gene. Thedifferentially expressed genes demonstrated at least a three-fold change in signal when comparing between tissue samples. The differences were all statistically significant (p<0.001) when compared to controls using a T-test.

3. Real Time PCR Analysis

Reverse transcription reactions were performed using up to 3.6 ug mRNA. The RNA was incubated for five minutes at 70.degree. C. and then chilled on ice. A master mix was added containing dNTPs, RT buffer (259 mM Tris-HCl pH 8.3, 375 mM KCl, 15mM MgCl.sub.2), dithiothreitol, random hexamers, RNasin, and reverse transcriptase (Life Technologies, Rockville, Md.). The reactions were incubated for 60 minutes at 37.degree. C., denatured for 5 minutes at 90.degree. C., then chilled on ice for 5minutes. PCR reactions were performed on ABI Prism.RTM. 5700 Sequence Detection System with SYBR green core reagents (PE Applied Biosystem, Foster City, Calif.). All PCR was done at 40 cycles with a pre-incubation period of 50.degree. C. for 2minutes followed by 95.degree. C. for 10 minutes. Cycling conditions were 95.degree. C. for 15 seconds, 55.degree. C. for 20 seconds, and 75.degree. C. for one minute. Some reactions were done with cycling conditions of 95.degree. C. for 15seconds and 60.degree. C. for 60 seconds. All data was normalized relative to the signal for the housekeeping gene human hypoxanthine phosphoribosyltransferase I ("HPRT") (Accession No. BC000578; GI: 12653602) (SEQ ID NO:2), the nucleotide sequence ofwhich is set forth in Table 2, below.

TABLE-US-00002 TABLE 2 Human Hypoxanthine Phosphoribosyltransferase I: Nucleotide Sequence Accession No. BC000578: GI: 12653602 (SEQ ID NO:2) 1 ggcacgaggc ctcctgagca gtcagcccgc gcgccggccg gctccgttat ggcgacccgc 61 agccctggcg tcgtgattag tgatgatgaaccaggttatg accttgattt attttgcata 121 cctaatcatt atgctgagga tttggaaagg gtgtttattc ctcatggact aattatggac 181 aggactgaac gtcttgctcg agatgtgatg aaggagatgg gaggccatca cattgtagcc 241 ctctgtgtgc tcaagggggg ctataaattc tttgctgacc tgctggatta catcaaagca 301ctgaatagaa atagtgatag atccattcct atgactgtag attttatcag actgaagagc 361 tattgtaatg accagtcaac aggggacata aaagtaattg gtggagatga tctctcaact 421 ttaactggaa agaatgtctt gattgtggaa gatataattg acactggcaa aacaatgcag 481 actttgcttt ccttggtcag gcagtataat ccaaagatggtcaaggtcgc aagcttgctg 541 gtgaaaagga ccccacgaag tgttggatat aagccagact ttgttggatt tgaaattcca 601 gacaagtttg ttgtaggata tgcccttgac tataatgaat acttcaggga tttgaatcat 661 gtttgtgtca ttagtgaaac tggaaaagca aaatacaaag cctaagatga gagttcaagt 721 tgagtttggaaacatctgga gtcctattga catcgccagt aaaattatca atgttctagt 781 tctgtggcca tctgcttagt agagcttttt gcatgtatct tctaagaatt ttatctgttt 841 tgtactttag aaatgtcagt tgctgcattc ctaaactgtt tatttgcact atgagcctat 901 agactatcag ttccctttgg gcggattgtt gtttaacttg taaatgaaaaaattctctta 961 aaccacagca ctattgagtg aaacattgaa ctcatatctg taagaaataa agagaagata 1021 tattagtttt ttaattggta ttttaatttt tatatatgca ggaaagaata gaagtgattg 1081 aatattgtta attataccac cgtgtgttag aaaagtaaga agcagtcaat tttcacatca 1141 aagacagcat ctaagaagttttgttctgtc ctggaattat tttagtagtg tttcagtaat 1201 gttgactgta ttttccaact tgttcaaatt attaccagtg aatctttgtc agcagttccc 1261 ttttaaatgc aaatcaataa attcccaaaa atttaaaaaa aaaaaaaaaa aaaaaa

Primer sets were as follows:

TABLE-US-00003 HPRT: Forward: GGTATACTGCCTGACCAAGG (SEQ ID NO:3) Reverse: CGAGATGTGATGAAGGAGATGG (SEQ ID NO:4) Name: gi475254 homo sapiens transcription factor ISGF-3 mRNA Forward exon 3 CCCCATGGAAATCAGACAGT (SEQ ID NO:5) Reverse exon 4TTGCTTTTCCGTATGTTGTG (SEQ ID NO:6) Name: gi28965 human alpha-1-antitrypsin gene (S variant) Forward TGAAGAGCGTCCTGGGTC (SEQ ID NO:7) Reverse CGTCGATGGTCAGCACAG (SEQ ID NO:8) Name: gi5595355 human ADO37 protein Forward GCCCATCAGTGACAGCAAG (SEQ ID NO:9)Reverse CCCAGGCAATGTTGAGGAG (SEQ ID NO:10) Name: gi2185828 human hypothetical protein FLI14834 Forward 417 436 CCTTCCCCTGTCATTGTTC Tm = 58 (SEQ ID NO:11) Reverse 515 534 GACAGTAACCCTGCCACAC Tm = 60 (SEQ ID NO:12) Name: gi183001 human guanylate bindingprotein isoform I Forward 124 132 GGCGACTGATGGCGAATC Tm = 58 (SEQ ID NO:13) Reverse 264 282 CACCGTGGAGCCCAGAGA Tm = 60 (SEQ ID NO:14) Name: gi2138110 human cysteine dioxygenease Forward 321 341 exon 1 GGCGATGAGGTCAATGTAGA Tm = 60 (SEQ ID NO:15) Reverse473 493 exon 2 CTGTGTCCTTCACCCCAACA Tm = 62 (SEQ ID NO:16) Name: gi180278 IgG Fc Receptor I Forward GGACACCACAAAGGCAGTGAT (SEQ ID NO:17) Reverse GCAGATGGAGCACCTCACAGT (SEQ ID NO:18) Name: gi1382379 MRP-14 Forward AGCTCAGCTGCTTGTCTGCAT (SEQ ID NO:19)Reverse TTCAAAGAGCTGGTGCGAAA (SEQ ID NO:20) Name: gi1382285 Early B Cell Factor Forward GGCCAGGGCAATGTTATGC (SEQ ID NO:21) Reverse ACATTCTGGCCCTCTGATCCT (SEQ ID NO:22) Name: gi2185828 human hypothetical protein FLI14834 Forward 417 436CCTTCCCCTGTCATTGTTC (SEQ ID NO:23) Reverse 515 534 GACAGTAACCCTGCCACAC (SEQ ID NO:24)

B. Further Characterization of GBP-1 (SEQ ID NOS. 41 and 42): and GBP-5 (SEQ ID NOS. 61 and 62) 1. Cell Culture

For real time PCR analyses, THP-1 cells (10.sup.7/group) were cultured at 10.sup.6/ml in RPMI containing 10% heat inactivated fetal calf serum, 2 mM L-glutamine with either medium, BMS-205820 (2 uM), or dexamethasone (100 nM) for 30 minutes at37.degree. C. in 5% CO.sub.2. LPS was added to each group (100 ng/ml), and the incubation continued for 0.5-8 hr. At the end of each time point, cells were pelleted, washed one time with 10 ml PBS, and stored at -80.degree. C.

Wild type 3T3 fibroblasts and immortalized fibroblast lines derived from p65 and IkB.alpha. germline knockouts were cultured in DMEM with 10% calf serum, glutamax and penicillin/streptomycin. Primary embryonic fibroblasts derived from germlineknockouts of relB and p50 were cultured in DMEM with 10% fetal calf serum, glutamax and penicillin/streptomycin. The fibroblasts were plated at 5.times.10.sup.5/well of a 6 well plate and cultured overnight at 37.degree. C. in 5% CO.sub.2. Cells werestimulated for 2 or 8 hours with either medium, human TNF.alpha. (10 ng/ml) or PMA (10 ng/ml). At each time point, cells were harvested using trypsin/EDTA, washed one time with PBS, and stored at -80.degree. C.

For the microarray analyses, THP-1 cells (10.sup.7/group) were cultured at 10.sup.6/ml as above for 1, 6, 24, or 48 hours with either medium, TNF.alpha. (10 ng/ml), IFN-.gamma. (100 U/ml), or LPS (100 ng/ml). At each time point, mRNA wasisolated, labeled, and used to probe Affymetrix HG_U95Av2, HG_U95B, and HG_U95C chips.

Human microvascular endothelial cells from three different donors were obtained from Clonetics (Walkersville, Md.), and cultured in EGM-2 medium (Clonetics). Cells were cultured in 100 mm dishes coated with mouse type IV collagen and allowed togrow to approximately 80% confluency. The cells were then stimulated for 1, 6, or 24 hours with either medium, TNF.alpha. (10 ng/ml), or IL-1.beta. (10 ng/ml). At each time point, mRNA was isolated, labeled and used to probe Affymetrix HG_U95Av2 andHG_U95B chips.

For VEGF and bFGF stimulations, microvascular endothelial cells from three independent donors were obtained, cultured in 100 mm dishes coated with mouse type IV collagen and allowed to grow to 30% confluency. At this time, the media was replacedwith DMEM containing 2% fetal calf serum, and the cells were cultured an additional day. The cells were stimulated for 1, 6, or 24 hours with either medium, VEGF (30 ng/ml) or bFGF (10 ng/ml). At each time point, mRNA was isolated, labeled, and used toprobe Affymetrix chips as described above.

Synovial fibroblasts were obtained from Cell Applications, Inc. (San Diego, Calif.), and cultured for 1, 6, or 24 hours with either medium, TNF.alpha. (10 ng/ml), IL-1.alpha. (10 ng/ml, Peprotech), IL-17 (10 ng/ml, R&D Systems, Minneapolis,Minn.), or IL-17b-Ig fusion protein (5 ng/ml). The IL-17b protein was produced by fusing the full length IL-17b sequence (Shi et al., J. Biol. Chem. 275:19167-19176 (2000)) to the human IgG1 Fc region. At each time point, mRNA was isolated, labeled,and used to probe Affymetrix HG_U95Av2, HG_U95B, HG_U95C, HG_U95D, and HG_U95E chips.

T cells were isolated from the blood of 4 donors. Lymphocytes were isolated by centrifugation over Accu-prep lymphocyte separation medium (Accurate Chemical and Scientific Corporation, Westbury, N.Y.). The T cells were isolated by rosettingwith sheep red blood cells. The isolated T cells were cultured for 6 hours with either medium, or immobilized anti-CD3 (1 ug/ml) plus soluble anti-CD28 (1 ug/ml) antibodies. After 6 hours, mRNA was isolated, labeled, and used to probe AffymetrixHG_U133A and HG_U133B chips.

2. cDNA Synthesis for Real Time PCR Analysis

Total RNA was isolated from cells using the RNeasy.RTM. Kit from Qiagen (Valencia, Calif.), including the on-the-column DNase digestion procedure. RNA quality and quantity were evaluated using UV spectrometry. Total RNA was used forfirst-strand cDNA synthesis using the SuperScript.TM. First-Strand Synthesis System for RT-PCR (Invitrogen, Carlsbad, Calif.) following the manufacturer's instructions with 50 ng of random hexamers.

For tissue expression analyses, Human Multiple Tissue cDNA Panel I and Human Immune System MTC Panels were obtained from Clontech (Palo Alto, Calif.). PCR reactions were performed using 2 microliters of cDNA sample (diluted with six microlitersof water).

3. Primers

Gene specific primers were designed using the Primer Express software and synthesized by Sigma Genosys (The Woodlands, Tex.).

Primer sets were as follows:

TABLE-US-00004 Name: mGBP-1 (SEQ ID NO:82) Forward GGAACAGGAAAGACTTCTCAAGCA (SEQ ID NO:83) Reverse CTTGACGTAGTTGCAAGCTCTCA Name: mGBP-5 (SEQ ID NO:84) Forward GCTGAAGCAAGGTAGCGATGA (SEQ ID NO:85) Reverse CCTCGTTGCTGAGTGTTGGA Name: mHPRT (SEQ IDNO:86) Forward TCAGACTGAAGAGCTACTGTAATGATCA (SEQ ID NO:87) Reverse CAACAATCAAGACATTCTTTCCAGTT Name: hGBP-5 (SEQ ID NO:88) Forward TGCTTTCACTTGTGCCTCTTTC (SEQ ID NO:89) Reverse CAGGCTCTCACAGAGACGGAA Name: hGAPDH (SEQ ID NO:90) Forward AGCCGAGCCACATCGCT(SEQ ID NO:91) Reverse GTGACCAGGCGCCCAATAC

3. PCR Assay Conditions

Reactions were performed in a total volume of 40 .mu.l. The master mix contained SYBR Green I Dye, 50 mM Tris-HCl pH 8.3, 75 mM KCl, DMSO, Rox reference dye, 5 mM MgCl.sub.2, 2 mM dNTP, Platinum Taq High Fidelity (1U/reaction), and 0.5 .mu.M ofeach primer. Eight microliters of diluted cDNA was used in each PCR reaction. The amplification program consisted of a 10 minute incubation at 95.degree. C. followed by forty cycles of incubations at 95.degree. C. for 15 seconds and 60.degree. C.for 1 minute. Amplification was followed by melting curve analysis at 60.degree. C. to demonstrate that the amplification was specific to a single amplicon. A negative control without cDNA template was run to assess the overall specificity.

4. Data Analysis

A relative value for the initial target concentration in each reaction was determined using the TaqMan 5700 software. The threshold value was set to 0.5 to obtain cycle threshold values that were used to assign relative message levels for eachtarget. The message levels of hGAPDH were used to normalize all other genes tested from the same cDNA. Message levels from the mouse fibroblast experiment were normalized using mouse HPRT values.

EXAMPLE 1

Upregulated Genes and Downregulated Genes in RA

1. .alpha.-1 Antitrypsin Expression

Using the materials and methods described hereinabove (Materials and Methods, Section A), increases in expression of .alpha.-1 antitrypsin were detected in the RA synovium. This increased expression is shown in the microarray data in FIG. 1. The polynucleotide sequence (SEQ ID NO:25) and amino acid sequence (SEQ ID NO:26) of .alpha.-1 antitrypsin are shown in FIGS. 2 and 3, respectively.

Using the materials and methods described hereinabove (Materials and Methods, Section A), Real Time PCR was conducted to quantify the expression of .alpha.-1 antitrypsin in the RA synovium, the results of which are set forth in Table 3, below. As used in Table 3 and hereinafter, "OA" stands for "Osteoarthritis".

TABLE-US-00005 TABLE 3 .alpha.-1 antitrypsin: Real Time PCR Results Expression Level T test Normal 1 OA 1.02 0.9574 RA 5.94 0.0034

.alpha.-1 Antitrypsin is the major endogenous inhibitor of the serine protease elastase. It also inhibits other circulating proteases including cathepsin G, thrombin, trypsin, and chymotrypsin. .alpha.-1 Antitrypsin is primarily synthesized bythe liver. However, neutrophils, monocytes, and alveolar macrophages also increase expression of .alpha.-1 antitrypsin in response to proinflammatory stimuli including TNF-.alpha., IL-6 and endotoxin (Knoell, et al., Am. J. Respir. Crit. Care Med. 157:246-255 (1998)). The deficiency of .alpha.-1 antitrypsin is associated with connective tissue destruction and the development of emphysema (Crystal, J. Clin. Invest. 85:1343-1352 (1990)).

At physiological concentrations, (.alpha.-1 antitrypsin is a potent stimulator of fibroblast proliferation and collagen production (Dabbagh et al., J. Cell Physiol. 186:73-81 (2001)). High levels of the elastase-.alpha.-1 antitrypsin complexhave been measured in the serum and synovial fluid of RA patients (Beyeler, et al., J. Rheumatol. 27:15-19 (2000)). .alpha.-1 Antitrypsin has also been isolated in a subtraction library examining genes upregulated in monocytes from RA patients(Stuhlmuller, et al., Arthritis Rheum. 43:775-790 (2000)).

2. B Lymphocyte Stimulator Expression

Using the materials and methods described hereinabove (Materials and Methods, Section A), increases in expression of B Lymphocyte Stimulator (BLyS, TNSF13B) were detected in the RA synovium. This increased expression is shown in the microarraydata in FIG. 4. The polynucleotide sequence (SEQ ID NO:27) and amino acid sequence (SEQ ID NO:28) of BLyS are shown in FIGS. 5 and 6, respectively.

BLyS is a member of the TNF family produced by activated T cells, monocytes, and dendritic cells that stimulates B cell expansion and function. Serum BLyS levels have been shown to be elevated in patients with systemic autoimmune diseases,including lupus erythematosus (Zhang, et al., J. Immunol. 166:6-10 (2001)) and RA (Cheema, et al., Arthritis Rheum. 44:1313-1319 (2001)). Mice deficient for the BLyS receptor are resistant to collagen-induced arthritis (Wang, et al., Nature Immunol. 2:632-637 (2001)).

3. Fc Gamma RI Expression

Using the materials and methods described hereinabove (Materials and Methods, Section A), increases in expression of Fc gamma RI were detected in the RA synovium. This increased expression is shown in the microarray data in FIG. 7. Thepolynucleotide sequence (SEQ ID NO:29) and amino acid sequence (SEQ ID NO:30) of Fc gamma RI are shown in FIGS. 8 and 9, respectively.

Using the materials and methods described hereinabove (Materials and Methods, Section A), Real Time PCR was conducted to quantify the expression of Fc gamma RI in the RA synovium, the results of which are set forth in Table 4, below.

TABLE-US-00006 TABLE 4 Fc gamma RI: Real Time PCR Results Expression Level T test Normal 1 1 OA 1.89 0.35 RA 6.57 0.02

Fc gamma RI receptors bind IgG immune complexes and trigger cell activation and IL-8 secretion (Salmon, et al., Arthritis Rheum. 44:739-750 (2001)). The expression of Fc gamma RI was increased on monocytes derived from RA patients as comparedto healthy controls. A significant correlation between Fc gamma RI, C-reactive protein, and blood platelet count was found in the RA patients. Furthermore, treatment with the steroid prednisolone induced down-regulation of Fc gamma RI expressionsuggesting that increased expression is associated with disease activity (Torsteinsdottir et al. (1999) Clin. Exp. Immunol. 115:554-560). Mice lacking functional Fc gamma RI and RIII receptors are resistant to collagen-induced arthritis (Kleinau etal. (2000) J. Exp. Med. 191:1611-1616).

4. Migration Inhibitory Factor-related Protein 14 Expression

Using the materials and methods described hereinabove (Materials and Methods, Section A), increases in expression of Migration inhibitory factor-related protein 14 (MRP-14) were detected in the RA synovium. This increased expression is shown inthe microarray data in FIG. 10. The polynucleotide sequence (SEQ ID NO:31) and amino acid sequence (SEQ ID NO:32) of MRP-14 are shown in FIGS. 11 and 12, respectively.

Using the materials and methods described hereinabove (Materials and Methods, Section A), Real Time PCR was conducted to quantify the expression of MRP-14 in the RA synovium, the results of which are set forth in Table 5, below.

TABLE-US-00007 TABLE 5 MRP-14: Real Time PCR Results Expression Level T test Normal 1 1 OA 0.64 0.49 RA 4.37 0.07

MRP-14 is a calcium binding protein expressed primarily by circulating neutrophils and monocytes that belongs to the S100 family of proteins (Kerkhoff, et al., Biochim. Biophys. Acta 1448:200-211 (1998); Hessian, et al., J. Leuk. Biol. 53:197-204 (1993)). MRP-14 is strongly expressed by infiltrating neutrophils and monocytes within the inflamed joints of juvenile RA patients (Youssef, et al., J. Rheumatol. 26:2523-2528 (1999)). MRP14 is specifically released during the interactionof monocytes with inflammatory activated endothelium, and is found in high concentrations in the synovial fluid of juvenile RA patients (Frosch, et al., Arthritis Rheum. 43:628-637 (2000)).

5. Skin Collagenase Expression

Using the materials and methods described hereinabove (Materials and Methods, Section A), increases in expression of Skin Collagenase (MMP-1) were detected in the RA synovium. This increased expression is shown in the microarray data in FIG. 13. The polynucleotide sequence (SEQ ID NO:33) and amino acid sequence (SEQ ID NO:34) of MRP-14 are shown in FIGS. 14 and 15, respectively.

MMP-1 is an enzyme that degrades interstitial collagens types I, II, and III. Elevated expression of MMP-1 was detected in synovium from patients with early inflammatory arthritis and with established erosive arthritis. Little expression wasdetected in normal synovium (Cunnane, et al., Rheumatology 38:34-42 (1999)). Primary cultures of rheumatoid synoviocytes produced MMP-1 as detected using immunohistochemistry. Expression has also been detected in the rheumatoid lesion (Tetlow, et al.,Br. J. Rheum. 37:64-70 (1998)).

6. Cysteine Dioxygenase Expression

Using the materials and methods described hereinabove (Materials and Methods, Section A), decreases in expression of cysteine dioxygenase were detected in the RA synovium. This decreased expression is shown in the microarray data in FIG. 16. The polynucleotide sequence (SEQ ID NO:35) and amino acid sequence (SEQ ID NO:36) of cysteine dioxygenase are shown in FIGS. 17 and 18, respectively.

Using the materials and methods described hereinabove (Materials and Methods, Section A), Real Time PCR was conducted to quantify the expression of Cysteine Dioxygenase in the RA synovium, the results of which are set forth in Table 6, below.

TABLE-US-00008 TABLE 6 Cysteine Dioxygenase: Real Time PCR Results Expression Level T test Normal 1 OA 1.21 0.77 RA 0.19 0.06

Cysteine dioxygenase is an enzyme involved in sulphate metabolism whose activity has been shown to be decreased in RA patients (Bradley, et al., J. Rheumatol. 21:1192-1196 (1994)).

EXAMPLE 2

Upregulated Genes and Downregulated Genes in RA

1. HLA-DR2/Dw12 Expression

Using the materials and methods described hereinabove (Materials and Methods, Section A), significant increases in expression of HLA-DR2/Dw12 were detected in the RA synovium. This increased expression is shown in the microarray data in FIG. 19. The polynucleotide sequence (SEQ ID NO:37) and amino acid sequence (SEQ ID NO:38) of HLA-DR2/Dw12 are shown in FIGS. 20 and 21, respectively.

Upregulation of MHC class II alleles, specifically HLA-DRB1 and HLA-DR4 subtypes, has previously been associated with development of RA (Kerlan-Candon, et al., Arthritis Rheum. 44:1281-1292 (2001)). Evidence suggests that expression of theDRB1*0401 and related haplotypes predisposes individuals to RA (Nepom, Adv. Immunol. 68:315-332 (1998)). This allele is also associated with the most severe form of RA leading to extraarticular manifestations (Weyand, et al., J. Clin. Invest. 89:2033-2039 (1992)). Specific associations of HLA-DR2 expression with RA, shown in the present invention, have not been previously demonstrated.

2. Stimulator of Iron Transport Expression

Using the materials and methods described hereinabove (Materials and Methods, Section A), increases in expression of Stimulator of Iron Transport were detected in the RA synovium. This increased expression is shown in the microarray data in FIG.22. The polynucleotide sequence (SEQ ID NO:39) and amino acid sequence (SEQ ID NO:40) of Stimulator of Iron Transport are shown in FIGS. 23 and 24, respectively.

Stimulator of Iron Transport is a regulator of ferric and ferrous iron uptake (Yu et al., J. Biol. Chem. 273:21380-21385 (1998); Gutierrez, et al., J. Cell Biol. 139:895-905 (1997)).

3. Guanylate Binding Protein Isoform 1 Expression

Using the materials and methods described hereinabove (Materials and Methods, Section A), increases in expression of Guanylate Binding Protein Isoform 1 (GBP-1) were detected in the RA synovium. This increased expression is shown in themicroarray data in FIG. 25. The polynucleotide sequence (SEQ ID NO:41) and amino acid sequence (SEQ ID NO:42) of GBP-1 are shown in FIGS. 26 and 27 respectively.

Using the materials and methods described hereinabove (Materials and Methods, Section A), Real Time PCR was conducted to quantify the expression of GBP-1 in the RA synovium, the results of which are set forth in Table 7, below.

TABLE-US-00009 TABLE 7 Guanylate Binding Protein Isoform 1: Real Time PCR Results Expression Level T test Normal 1 OA 1.00 0.985 RA 3.89 0.003

GBP-1 is an interferon-inducible protein that binds guanine nucleotides and possesses GTPase activity (Cheng, et al., Mol. Cell. Biol. 11:4717-4725 (1991)).

The regulation of GBP-1 was further characterized using the materials and methods described hereinabove (Materials and Methods, Section B), the results of which are set forth in Example 3 below.

4. ISGF-3 p91 (STAT1) Expression

Using the materials and methods described hereinabove (Materials and Methods, Section A), increases in expression of ISGF-3 p91 (STAT1) were detected in the RA synovium. This increased expression is shown in the microarray data in FIG. 28. Thepolynucleotide sequence (SEQ ID NO:43) and amino acid sequence (SEQ ID NO:44) of ISGF-3 p91 (STAT1) are shown in FIGS. 29 and 30, respectively.

Using the materials and methods described hereinabove (Materials and Methods, Section A), Real Time PCR was conducted to quantify the expression of ISGF-3 p91 in the RA synovium, the results of which are set forth in Table 8, below.

TABLE-US-00010 TABLE 8 ISGF-3 p91: Real Time PCR Results Expression Level T test Normal 1 OA 0.93 0.720 RA 3.91 0.002

ISGF-3 p91 (STAT1) is a transcription factor involved in interferon signaling pathways (Schindler, et al., Proc. Nat. Acad. Sci. 89:7836-7839 (1992)). Continuous activation of STAT1 has been seen in synovial fluid cells derived from RA butnot osteoarthritis patients (Yokota, et al., J. Rheumatol. 28:1952-1959 (2001)).

5. Mad Protein Homolog-3 Expression

Using the materials and methods described hereinabove (Materials and Methods, Section A), decreases in expression of Mad Protein Homolog (MAD-3) were detected in the RA synovium. MAD-3 is also known as Mothers Against Decapentaplegic Homolog 3(Smad-3). This decreased expression is shown in the microarray data in FIG. 31. The polynucleotide sequence (SEQ ID NO:45) and amino acid sequence (SEQ ID NO:46) of MAD-3 are shown in FIGS. 32 and 33, respectively.

Mad-3 is an intracellular mediator downstream of the TGF-.beta./activin receptors that appears to be important for monocyte chemotaxis in response to TGF-.beta. (Zhang, et al., Nature 383:168-172 (1996); Ashcroft, et al., Nature Cell Biol. 1:260-266 (1999).

6. Human Transforming Growth Factor-Beta Type III Receptor Expression

Using the materials and methods described hereinabove (Materials and Methods, Section A), decreases in expression of Human Transforming Growth Factor-Beta Type III Receptor (TGF-.beta. type III receptor) were detected in the RA synovium. Thisdecreased expression is shown in the microarray data in FIG. 34. The polynucleotide sequence (SEQ ID NO:47) and amino acid sequence (SEQ ID NO:48) of TGF-.beta. type III receptor are shown in FIGS. 35 and 36, respectively.

7. Early B Cell Factor Expression

Using the materials and methods described hereinabove (Materials and Methods, Section A), decreases in expression of Early B Cell Factor (EBF) were detected in the RA synovium. This decreased expression is shown in the microarray data in FIG.37. The polynucleotide sequence (SEQ ID NO:49) and amino acid sequence (SEQ ID NO:50) of EBF are shown in FIGS. 38 and 39, respectively.

Using the materials and methods described hereinabove (Materials and Methods, Section A), Real Time PCR was conducted to quantify the expression of EBF in the RA synovium, the results of which are set forth in Table 9, below.

TABLE-US-00011 TABLE 9 EBF: Real Time PCR Results Expression Level T test Normal 1 1 OA 0.72 0.66 RA 0.20 0.05

EBF is a transcription factor required for B cell differentiation (Gisler, et al., Blood 96:1457-1464 (2000)).

8. Duodenal Cytochrome b Expression

Using the materials and methods described hereinabove (Materials and Methods, Section A), decreases in expression of Duodenal Cytochrome b were detected in the RA synovium. This decreased expression is shown in the microarray data in FIG. 40. The polynucleotide sequence (SEQ ID NO:51) and amino acid sequence (SEQ ID NO:52) of Duodenal Cytochrome b are shown in FIGS. 41 and 42, respectively.

Duodenal cytochrome b is a protein localized to the duodenal mucosa possessing ferric reductase activity (McKie, et al., Science 291:1755-1759 (2001)).

9. Nuclear LIM Interactor-Interacting Factor Expression

Using the materials and methods described hereinabove (Materials and Methods, Section A), decreases in expression of Nuclear LIM Interactor-Interacting Factor (NLI-IF) were detected in the RA synovium. This decreased expression is shown in themicroarray data in FIG. 43. The polynucleotide sequence (SEQ ID NO:53) and amino acid sequence (SEQ ID NO:54) of NLI-IF are shown in FIGS. 44 and 45, respectively.

The NLI-IF amino acid sequence has homology to the nuclear Lim interactor interacting factor from Gallus gallus. It is one of a family of four related proteins of unknown function (Marquet, et al., Mamm. Genome 11:755-762 (2000)).

10. Deleted in Liver Cancer 1 Expression

Using the materials and methods described hereinabove (Materials and Methods, Section A), decreases in expression of Deleted in Liver Cancer 1 (DLC1) were detected in the RA synovium. This decreased expression is shown in the microarray data inFIG. 46. The polynucleotide sequence (SEQ ID NO:55) and amino acid sequence (SEQ ID NO:56) of DLC1 are shown in FIGS. 47 and 48, respectively.

DLC1 is a candidate tumor suppressor gene possessing a high degree of sequence similarity to the rat p122 RhoGap gene (Yuan, et al., Cancer Res. 58:2196-2199 (1998); Ng, et al., Cancer Res. 60:6581-6584 (2000)).

11. GI: 12005907

Using the materials and methods described hereinabove (Materials and Methods, Section A), increases in expression of the polynucleotide identified by Genbank Accession No. AF260335 (GI: 12005907) were detected in the RA synovium. This increasedexpression is shown in the microarray data in FIG. 49. The polynucleotide sequence (SEQ ID NO:57) and amino acid sequence (SEQ ID NO:58) are shown in FIGS. 50 and 51, respectively.

Further, Real Time PCR was conducted to quantify the expression of this polynucleotide in the RA synovium, the results of which are set forth in Table 10.

TABLE-US-00012 TABLE 10 GI: 12005907: Real Time PCR Results Expression Level T test Normal 1 OA 1.44 0.34 RA 2.39 0.04

12. Apolipoprotein L Expression

Using the materials and methods described hereinabove (Materials and Methods, Section A), increases in expression of Apolipoprotein L (APOL) (Genbank Accession No. NM.sub.--003661) were detected in the RA synovium. This increased expression isshown in the microarray data in FIG. 52. The polynucleotide sequence (SEQ ID NO:59) and amino acid sequence (SEQ ID NO:60) of APOL are shown in FIGS. 53 and 54, respectively.

Apolipoprotein L is a component of human plasma lipoproteins (Duchateau, et al., J. Biol. Chem. 272:25576-25582 (1997)).

13. Homo Sapiens Guanylate Binding Protein 5 Expression

Using the materials and methods described hereinabove (Materials and Methods, Section A), increases in expression of Homo Sapiens Guanylate Binding Protein 5 (Genbank Accession No. AF288815) (GBP-5) were detected in the RA synovium. Thisincreased expression is shown in the microarray data in FIG. 55. The polynucleotide sequence (SEQ ID NO:61) and amino acid sequence (SEQ ID NO:62) of Homo Sapiens GBP-5 are shown in FIGS. 56 and 57, respectively. GBP-5 is highly homologous to GBP-1described above (SEQ ID NO: 42).

The regulation of GBP-5 was further characterized using the materials and methods described hereinabove (Materials and Methods, Section B), the results of which are set forth in Example 3 below.

14. Human Proteasome Activator hPA28 Subunit Beta Expression

Using the materials and methods described hereinabove (Materials and Methods, Section A), increases in expression of HPA28 subunit beta (HPA28) (Genbank Accession No. D45248) were detected in the RA synovium. This increased expression is shownin the microarray data in FIG. 58. The polynucleotide sequence (SEQ ID NO:63) and amino acid sequence (SEQ ID NO:64) of HPA28 are shown in FIGS. 59 and 60, respectively.

HPA28 beta subunits associate with alpha subunits to form PA28, an activator of the 20S proteasome. Both subunits are coordinately regulated by interferon-.gamma. (Ahn, et al., FEBS Lett. 366:37-42 (1995)).

15. Homo Sapiens FYN Binding Protein

Using the materials and methods described hereinabove (Materials and Methods, Section A), increases in expression of Homo Sapiens FYN Binding Protein (Genbank Accession No. AF001862) were detected in the RA synovium. This increased expression isshown in the microarray data in FIG. 61. The polynucleotide sequence (SEQ ID NO:65) and amino acid sequence (SEQ ID NO:66) of Homo Sapiens FYN Binding Protein are shown in FIGS. 62 and 63, respectively.

FYN Binding Protein is a hematopoietic specific adapter protein that associates in a T cell receptor-inducible manner with another hematopoietic-specific adapter, SLP-76 (daSilva, et al., Proc. Natl. Acad. Sci. USA 94:7493-7498 (1997)). Tcells from mice lacking FYN Binding Protein exhibit impaired proliferative responses and impaired integrin clustering following T cell receptor crosslinking (Peterson, et al., Science 293:2263-2265 (2001)).

16. VAMP5

Using the materials and methods described hereinabove (Materials and Methods, Section A), increases in expression of VAMP5 (GI:4027902) were detected in the RA synovium. This increased expression is shown in the microarray data in FIG. 64. Thepolynucleotide sequence (SEQ ID NO:67) and amino acid sequence (SEQ ID NO:68) of VAMP5 are shown in FIGS. 65 and 66, respectively.

VAMP5 is a novel synaptobrevin protein that is preferentially expressed in skeletal muscle and heart. Its expression is increased during myogenesis and it localizes to the plasma membrane as well as intracellular perinuclear and peripheralvesicular structures of myotubes (Zeng, et al., Mol. Biol. Cell 9:2423-2437 (1998)).

17. GI: 2466183

Using the materials and methods described hereinabove (Materials and Methods, Section A), decreases in expression of the polynucleotide identified by GI:2466183 were detected in the RA synovium. This decreased expression is shown in themicroarray data in FIG. 67. The sequence of this polynucleotide is shown in FIG. 68 (SEQ ID NO:69).

18. GI: 22192831

Using the materials and methods described hereinabove (Materials and Methods, Section A), decreases in expression of the polynucleotide identified by GI: 2219283 were detected in the RA synovium. This decreased expression is shown in themicroarray data in FIG. 69. The sequence of this polynucleotide is shown in FIG. 70 (SEQ ID NO:70).

19. Hypothetical Protein FLJ20152 (GI: 9506660)

Using the materials and methods described hereinabove (Materials and Methods, Section A), decreases in expression of the Hypothetical Protein FLJ20152 identified by GI: 9506660 were detected in the RA synovium. This decreased expression is shownin the microarray data in FIG. 71. The polynucleotide sequence (SEQ ID NO:71) and amino acid sequence (SEQ ID NO:72) of Hypothetical Protein FLJ20152 are shown in FIGS. 72 and 73, respectively.

20. GI: 5876137

Using the materials and methods described hereinabove (Materials and Methods, Section A), decreases in expression of the polynucleotide identified by GI: 5876137 were detected in the RA synovium. This decreased expression is shown in themicroarray data in FIG. 74. The sequence of this polynucleotide is shown in FIG. 75 (SEQ ID NO:73).

21. GI: 2185828

Using the materials and methods described hereinabove (Materials and Methods, Section A), decreases in expression of the polynucleotide identified by GI: 2185828 were detected in the RA synovium. This decreased expression is shown in themicroarray data in FIG. 76. The polynucleotide sequence (SEQ ID NO:74) and amino acid sequence (SEQ ID NO:75) are shown in FIGS. 77 and 78, respectively.

Further, Real Time PCR was conducted to quantify the expression of this polynucleotide in the RA synovium, the results of which are set forth in Table 11.

TABLE-US-00013 TABLE 11 GI: 2185828 Real Time PCR Results Expression Level T test Normal 1 OA 0.87 0.632 RA 0.21 0.014

22. Homo Sapiens Proteasome (Prosome, Macropain) Subunit, Beta Type, 9

Using the materials and methods described hereinabove (Materials and Methods, Section A), increases in expression of the Homo sapiens proteasome (prosome, macropain) subunit, beta type, 9 (GI: 14754802) were detected in the RA synovium. Thisincreased expression is shown in the microarray data in FIG. 79. The polynucleotide sequence (SEQ ID NO:76) and amino acid sequence (SEQ ID NO:77) of homo sapiens proteasome (prosome, macropain) subunit, beta type, 9 are shown in FIGS. 80 and 81,respectively.

The of homo sapiens proteasome (prosome, macropain) subunit, beta type, 9 is encoded by a gene within the major histocompatibility complex. This subunit replaces beta subunit PSMB6 following interferon gamma stimulation, thereby altering theproteasome specificity.

23. TYRO Protein Tyrosine Kinase Binding Protein (TYROBP); GI:4507754

Using the materials and methods described hereinabove (Materials and Methods, Section A), increases in expression of the TYRO protein tyrosine kinase binding protein (TYROBP) (GI:4507754) were detected in the RA synovium. This increasedexpression is shown in the microarray data in FIG. 82. The polynucleotide sequence (SEQ ID NO:78) and amino acid sequence (SEQ ID NO:79) are shown in FIGS. 83 and 84, respectively.

TYRO protein tyrosine binding protein (TYROBP) is an ITAM-bearing transmembrane adaptor protein that associates non-covalently with receptors in natural killer and myeloid cells (Lanier, et al., Nature 391:703-707 (1998)). Mice deficient forTYROBP have normal lymphoid and myeloid development, however activating Ly49 receptors on NK cells are downregulated and nonfunctional. The TYROBP deficient mice are resistant to induction of experimental autoimmune encephalomyelitis and exhibitdecreased interferon-.gamma. production by antigen-primed CD4+ T cells due to inadequate T cell priming in vivo (Bakker, et al., Immunity 13:345-353 (2000)). Humans expressing loss of function mutations in TYROBP exhibit presenile dementia with bonecysts (Paloneva, et al., Nat. Genet. 25:357-361 (2000)).

24. Interleukin 15 Receptor, Alpha

Using the materials and methods described hereinabove (Materials and Methods, Section A), increases in expression of Interleukin 15 Receptor, alpha were detected in the RA synovium. This increased expression is shown in the microarray data inFIG. 85. The polynucleotide sequence (SEQ ID NO:80) and amino acid sequence (SEQ ID NO:81) are shown in FIGS. 86 and 87, respectively.

IL-15 is a T cell growth factor that shares many functional similarities with IL-2. The IL-15 receptor consists of a high affinity binding alpha chain and the common IL-2 receptor beta and gamma chains (Anderson, et al., J. Biol. Chem.270:29862-29869 (1995)). Elevated levels of IL-15 have been detected in the serum from systemic lupus erythematosus patients (Aringer, et al., Rheumatology 40:876-881 (2001)), in the synovial tissue of rheumatoid arthritis patients (Thurkow, et al., J.Pathol. 181:444-450 (1997)), and in synovial fluid from rheumatoid arthritis patients (McInnes, et al., Nat. Med. 2:175-182 (1996)). Administration of soluble IL-15 receptor alpha chain to mice prevented collagen-induced arthritis (Ruchatz, et al.,J. Immunol. 160:5654-5660 (1998)), suggesting that IL-15 plays a role in the development of rheumatoid arthritis. Upregulation of the IL-15 receptor alpha chain in rheumatoid arthritis has not been previously described.

EXAMPLE 3

Characterization of RA-Associated Genes GBP-1 and GBP5

As stated above, GBP-1 and GBP-5 were further characterized using the materials and methods set forth above (Materials and Methods, Section B).

GBP-1 has been identified as an interferon-inducible protein in human fibroblasts (Cheng et al., J. Biol. Chem. 258:7746-7750 (1983)). GBP-1 mRNA has been shown to be induced in endothelial cells in response to the pro-inflammatory stimuli,TNF.alpha. and IL-1.alpha. (Guenzi et al., EMBO J. 20:5568-5577 (2001)). This study also suggested that GBP-1 mediates the anti-proliferative effects of these cytokines.

As TNF.alpha. and IL-1.alpha. are known to activate the transcription factor NF-kB, it was determined whether NF-kB was required for induction of GBP-1 and GBP-5 in the human THP-1 monocyte line. THP-1 monocytes were stimulated withlipopolysaccharide (LPS), a known inducer of NF-kB, in the presence and absence of a selective peptide inhibitor of NF-kB nuclear translocation, which is set forth in Fujihara et al., J. Immunol. 165:1004-1012 (2000) and designated "BMS-205820".

Following a 2 hour stimulation, RNA was isolated from 2 sets of independently treated THPs, and real time PCR was performed using primers specific for either GBP-1 or GBP-5, as shown in FIG. 88. Treatment of THP-1 cells with LPS significantlyincreased steady-state mRNA levels of both GBP-1 (FIGS. 88A and 88B) and GBP-5 (FIGS. 88C and 88D). Expression of both genes was significantly inhibited by BMS-205820, suggesting that LPS-mediated induction of GBP-1 and GBP-5 expression is dependent onNF-kB activity.

An extended time course was performed to further characterize GBP-1 and GBP-5 expression, as shown in FIG. 89. THP-1 cells were stimulated for 0.5, 1, 4, 6, and 8 hours with LPS. Some groups included either BMS-205820 or the steroiddexamethasone for 2 or 6 hours. Dexamethasone is also known to inhibit NF-KB activity (Scheinman et al., Mol. Cell. Biol. 15:943-953 (1995)). At each time point, mRNA was isolated and real time PCR was performed using primers specific for eitherGBP-1 FIG. 89A) or GBP-5 (FIG. 89B). Steady state mRNA levels for both GBP-1 and GBP-5 peaked at 6 hours post stimulation. Addition of either BMS-205820 or dexamethasone significantly inhibited mRNA induction of both genes at 2 and 6 hours. Theability of two different NF-kB inhibitors to block GBP-1 and GBP-5 expression further confirms that LPS-mediated induction of these genes is dependent on NF-kB activity.

To further confirm that GBP-1 and GBP-5 are NF-kB target genes, expression in mouse embryonic fibroblasts derived from germline knockouts of members of the NF-kB family was examined (FIG. 90). Wild type 3T3 cells, embryonic fibroblasts derivedfrom knockouts of p65, RelB, p50, and IkB.alpha. were stimulated for 2 or 8 hours with either TNF.alpha. or PMA. At each time point, mRNA was isolated and real time PCR was performed using primers specific for either mouse GBP-1 (FIG. 90A) or GBP-5(FIG. 90B). Stimulation with TNF(.alpha. but not PMA induced increased steady-state levels of both GBP-1 and GBP-5 mRNA. Induction of GBP-1 mRNA was completely ablated in cells lacking either p65 or RelB. GBP-1 mRNA was superinduced in cells lackingeither p50 or IkB.alpha., suggesting that these proteins negatively regulate GBP-1 mRNA. IkB.alpha. is a known inhibitor of NF-kB activity (Baeuerle et al., Science 242:540-545 (1988)). Homodimers of p50 have also been shown to repress certain genes(Plaksin et al., J. Exp. Med. 177:1651-1662 (1993)).

Similar to GBP-1, induction of GBP-5 mRNA was completely ablated in cells lacking p65. In contrast to GBP-1, GBP-5 mRNA was superinduced in cells lacking RelB. Similar to GBP-1, GBP-5 mRNA was also superinduced in cells lacking either p50 orIkB.alpha.. These data suggest that p65 expression is required for the induction of both GBP-1 and GBP-5. Complexes containing RelB appear to differentially regulate GBP-1 and GBP-5 expression. Taken together, these data are consistent withNF-kB-dependent regulation of GBP-1 and GBP-5 expression.

The tissue expression profiles of GBP-1 and GBP-5 were further characterized. Human tissue cDNA panels were analyzed by real time PCR with primers selective for GBP-1 (FIG. 91A) and GBP-5 (FIG. 91B). Both genes had very similar patterns ofexpression. The highest steady state mRNA levels were detected in hematopoietic tissues including spleen, peripheral blood leukocytes, and lymph nodes. Lower levels of expression were detected in lung, followed by liver, thymus, tonsil, bone marrow,placenta, fetal liver, tonsil, and pancreas.

Based on the high expression detected in hematopoietic tissue, the expression of GBP-1 and GBP-5 in panels of resting and stimulated immune cells was examined. Consistent with the identification of GBP-1 as an interferon response gene (Cheng etal., J. Biol. Chem. 258:7746-7750 (1983)), steady state levels of GBP-1 and GBP-5 mRNA were strongly induced by interferon-.gamma. treatment of THP-1 monocytes, as shown in FIGS. 92A and 92B. Much lower levels of expression were induced by TNF.alpha. and LPS.

Consistent with published reports (Guenzi et al., EMBO J. 20:5568-5577 (2001)), GBP-1 expression was strongly induced by TNF.alpha. and IL-1.beta. in human microvascular endothelial cells (FIG. 93A). No induction was seen in response to eitherVEGF or basic FGF. In contrast to GBP-1, induction of GBP-5 by TNF.alpha. and IL-1.beta. was variable (FIG. 93B). Cells from two out of three donors upregulated GBP-5 mRNA in response to TNF.alpha.. Only one donor significantly induced GBP-5 mRNA inresponse to IL-1.beta..

Synovial fibroblasts derived from rheumatoid arthritis patients were stimulated with either TNF.alpha., IL-1.alpha., IL-17, or IL-17b. GBP-1 mRNA was induced at 1 and 6 hours by stimulation with either TNF.alpha. or IL-1.alpha., but not inresponse to either IL-17 or IL-17b (FIG. 94A). Low levels of GBP-5 expression were detected in synovial fibroblasts (FIG. 94B). Induction of GBP-5 in response to the different stimuli was variable and not sustained.

GBP-1 and GBP-5 had similar patterns of expression in T cells (FIG. 95). Peripheral blood T cells were isolated from 4 different donors and stimulated for 6 hours with antibodies to CD3 and CD28 as a mimic of antigen stimulation. Steady statelevels of GBP-1 (FIG. 95A) and GBP-5 (FIG. 95B) mRNA were strongly induced by antigen receptor crosslinking.

The induction of GBP-1 and GBP-5 by pro-inflammatory stimuli including LPS, IL-1, TNF.alpha., and antigen receptor crosslinking is consistent with NF-kB-dependent regulation of these genes. Overexpression of these genes in synovium fromrheumatoid arthritis patients is also consistent with NF-kB-dependent regulation. NF-kB is activated in the inflamed synovium of rheumatoid arthritis patients (Marok et al., Arthritis Rheum. 39:583-591 (1996)) and animal models of arthritis (Miagkov etal., Proc. Natl. Acad. Sci. USA 95:13859-13864 (1998)). The regulation of GBP-1 and GBP-5 by NF-kB coupled with the involvement of NF-kB in the development of arthritis indicates that these genes play a role in disease pathology.

While the invention has been described in connection with specific embodiments therefore, it will be understood that it is capable of further modifications and this application is intended to cover any variations, uses, or adaptations of theinvention following, in general, the principles of the invention and including such departures from the present disclosure as come within known or customary practice within the art to which the invention pertains and as may be applied to the essentialfeatures hereinbefore set forth and as follows in the scope of the appended claims. All references cited herein are expressly incorporated in their entirety.

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9Artificial SequenceArtificial Sequence equals primertgaa ttgtaatacg actcactata gggaggcggt tttttttttt tttttttttt 62homo sapiens 2ggcacgaggc ctcctgagca gtcagcccgc gcgccggccg gctccgttat ggcgacccgc 6ggcg tcgtgattag tgatgatgaa ccaggttatg accttgattt attttgcata atcattatgctgagga tttggaaagg gtgtttattc ctcatggact aattatggac ctgaac gtcttgctcg agatgtgatg aaggagatgg gaggccatca cattgtagcc 24gtgc tcaagggggg ctataaattc tttgctgacc tgctggatta catcaaagca 3tagaa atagtgatag atccattcct atgactgtag attttatcagactgaagagc 36aatg accagtcaac aggggacata aaagtaattg gtggagatga tctctcaact 42ggaa agaatgtctt gattgtggaa gatataattg acactggcaa aacaatgcag 48cttt ccttggtcag gcagtataat ccaaagatgg tcaaggtcgc aagcttgctg 54agga ccccacgaag tgttggatataagccagact ttgttggatt tgaaattcca 6gtttg ttgtaggata tgcccttgac tataatgaat acttcaggga tttgaatcat 66gtca ttagtgaaac tggaaaagca aaatacaaag cctaagatga gagttcaagt 72tgga aacatctgga gtcctattga catcgccagt aaaattatca atgttctagt 78gccatctgcttagt agagcttttt gcatgtatct tctaagaatt ttatctgttt 84ttag aaatgtcagt tgctgcattc ctaaactgtt tatttgcact atgagcctat 9atcag ttccctttgg gcggattgtt gtttaacttg taaatgaaaa aattctctta 96agca ctattgagtg aaacattgaa ctcatatctg taagaaataaagagaagata tagtttt ttaattggta ttttaatttt tatatatgca ggaaagaata gaagtgattg attgtta attataccac cgtgtgttag aaaagtaaga agcagtcaat tttcacatca acagcat ctaagaagtt ttgttctgtc ctggaattat tttagtagtg tttcagtaat gactgta ttttccaacttgttcaaatt attaccagtg aatctttgtc agcagttccc taaatgc aaatcaataa attcccaaaa atttaaaaaa aaaaaaaaaa aaaaaa DNAArtificial SequenceArtificial Sequence equals primer 3ggtatactgc ctgaccaagg 2Artificial SequenceArtificial Sequence equalsprimer. 4cgagatgtga tgaaggagat gg 2252ificial SequenceArtificial Sequence equals primer. 5ccccatggaa atcagacagt 2Artificial SequenceArtificial Sequence equals primer. 6ttgcttttcc gtatgttgtg 2Artificial SequenceArtificial Sequenceequals primer. 7tgaagagcgt cctgggtc AArtificial SequenceArtificial Sequence equals primer. 8cgtcgatggt cagcacag AArtificial SequenceArtificial Sequence equals primer. 9gcccatcagt gacagcaag NAArtificial SequenceArtificial Sequenceequals primer. gcaat gttgaggag NAArtificial SequenceArtificial Sequence equals primer. ccctg tcattgttc NAArtificial SequenceArtificial Sequence equals primer. taacc ctgccacac NAArtificial SequenceArtificialSequence equals primer. ctgat ggcgaatc NAArtificial SequenceArtificial Sequence equals primer. tggag cccagaga NAArtificial SequenceArtificial Sequence equals primer. tgagg tcaatgtaga 2AArtificialSequenceArtificial Sequence equals primer. tcctt caccccaaca 2AArtificial SequenceArtificial Sequence equals primer. ccaca aaggcagtga t 2AArtificial SequenceArtificial Sequence equals primer. tggag cacctcacag t2AArtificial SequenceArtificial Sequence equals primer. agctg cttgtctgca t 2AArtificial SequenceArtificial Sequence equals primer. 2gagc tggtgcgaaa 2AArtificial SequenceArtificial Sequence equals primer. 2ggcaatgttatgc NAArtificial SequenceArtificial Sequence equals primer. 22acattctggc cctctgatcc t 2AArtificial SequenceArtificial Sequence equals primer. 23ccttcccctg tcattgttc NAArtificial SequenceArtificial Sequence equals primer. 24gacagtaacc ctgccacac o sapiens 25ctgcaggggg gggggggggc tgggacagtg aatcgacaat gccgtcttct gtctcgtggg 6tcct gctggcaggc ctgtgctgcc tggtccctgt ctccctggct gaggatcccc agatgc tgcccagaag acagatacat cccaccatga tcaggatcac ccaaccttcagatcac ccccaacctg gctgagttcg ccttcagcct ataccgccag ctggcacacc 24acag caccaatatc ttcttctccc cagtgagcat cgctacagcc tttgcaatgc 3ctggg gaccaaggct gacactcacg atgaaatcct ggagggcctg aatttcaacc 36agat tccggaggct cagatccatg aaggcttccaggaactcctc cgtaccctca 42caga cagccagctc cagctgacca ccggcaatgg cctgttcctc agcgagggcc 48tagt ggataagttt ttggaggatg ttaaaaagtt gtaccactca gaagccttca 54actt cggggacacc gaagaggcca agaaacagat caacgattac gtggagaagg 6caagg gaaaattgtggatttggtca aggagcttga cagagacaca gtttttgctc 66atta catcttcttt aaaggcaaat gggagagacc ctttgaagtc aaggacaccg 72agga cttccacgtg gaccaggtga ccaccgtgaa ggtgcctatg atgaagcgtt 78tgtt taacatccag cactgtaaga agctgtccag ctgggtgctg ctgatgaaat84gcaa tgccaccgcc atcttcttcc tgcctgatga ggggaaacta cagcacctgg 9gaact cacccacgat atcatcacca agttcctgga aaatgaagac agaaggtctg 96taca tttacccaaa ctgtccatta ctggaaccta tgatctgaag agcgtcctgg aactggg catcactaag gtcttcagca atggggctgacctctccggg gtcacagagg cacccct gaagctctcc aaggccgtgc ataaggctgt gctgaccatc gacgagaaag ctgaagc tgctggggcc atgtttttag aggccatacc catgtctatc ccccccgagg agttcaa caaacccttt gtcttcttaa tgattgaaca aaataccaag tctcccctct tgggaaaagtggtgaat cccacccaaa aataactgcc tctcgctcct caacccctcc ccatccc tggccccctc cctggatgac attaaagaag ggttgagctg g mo sapiens 26Met Pro Ser Ser Val Ser Trp Gly Ile Leu Leu Ala Gly Leu Cys Cysal Pro Val Ser Leu Ala Glu Asp ProGln Gly Asp Ala Ala Gln 2Lys Thr Asp Thr Ser His His Asp Gln Asp His Pro Thr Phe Asn Lys 35 4 Thr Pro Asn Leu Ala Glu Phe Ala Phe Ser Leu Tyr Arg Gln Leu 5Ala His Gln Ser Asn Ser Thr Asn Ile Phe Phe Ser Pro Val Ser Ile65 7AlaThr Ala Phe Ala Met Leu Ser Leu Gly Thr Lys Ala Asp Thr His 85 9 Glu Ile Leu Glu Gly Leu Asn Phe Asn Leu Thr Glu Ile Pro Glu Gln Ile His Glu Gly Phe Gln Glu Leu Leu Arg Thr Leu Asn Gln Asp Ser Gln Leu Gln Leu Thr ThrGly Asn Gly Leu Phe Leu Ser Gly Leu Lys Leu Val Asp Lys Phe Leu Glu Asp Val Lys Lys Leu Tyr His Ser Glu Ala Phe Thr Val Asn Phe Gly Asp His Glu Glu Ala Lys Gln Ile Asn Asp Tyr Val Glu Lys Gly Thr Gln Gly LysIle Asp Leu Val Lys Glu Leu Asp Arg Asp Thr Val Phe Ala Leu Val 2yr Ile Phe Phe Lys Gly Lys Trp Glu Arg Pro Phe Glu Val Lys 222r Glu Asp Glu Asp Phe His Val Asp Gln Val Thr Thr Val Lys225 234o MetMet Lys Arg Leu Gly Met Phe Asn Ile Gln His Cys Lys 245 25s Leu Ser Ser Trp Val Leu Leu Met Lys Tyr Leu Gly Asn Ala Thr 267e Phe Phe Leu Pro Asp Glu Gly Lys Leu Gln His Leu Glu Asn 275 28u Leu Thr His Asp Ile Ile Thr Lys PheLeu Glu Asn Glu Asp Arg 29er Ala Ser Leu His Leu Pro Lys Leu Ser Ile Thr Gly Thr Tyr33sp Leu Lys Ser Val Leu Gly Gln Leu Gly Ile Thr Lys Val Phe Ser 325 33n Gly Ala Asp Leu Ser Gly Val Thr Glu Glu Ala Pro Leu Lys Leu345s Ala Val His Lys Ala Val Leu Thr Ile Asp Glu Lys Gly Thr 355 36u Ala Ala Gly Ala Met Phe Leu Glu Ala Ile Pro Met Ser Ile Pro 378u Val Lys Phe Asn Lys Pro Phe Val Phe Leu Met Ile Glu Gln385 39hr Lys SerPro Leu Phe Met Gly Lys Val Val Asn Pro Thr Gln 44858DNAHomo Sapiens 27atggatgact ccacagaaag ggagcagtca cgccttactt cttgccttaa gaaaagagaa 6aaac tgaaggagtg tgtttccatc ctcccacgga aggaaagccc ctctgtccga ccaaag acggaaagct gctggctgcaaccttgctgc tggcactgct gtcttgctgc cggtgg tgtctttcta ccaggtggcc gccctgcaag gggacctggc cagcctccgg 24ctgc agggccacca cgcggagaag ctgccagcag gagcaggagc ccccaaggcc 3ggagg aagctccagc tgtcaccgcg ggactgaaaa tctttgaacc accagctcca 36ggcaactccagtca gaacagcaga aataagcgtg ccgttcaggg tccagaagaa 42actc aagactgctt gcaactgatt gcagacagtg aaacaccaac tatacaaaaa 48taca catttgttcc atggcttctc agctttaaaa ggggaagtgc cctagaagaa 54aata aaatattggt caaagaaact ggttactttt ttatatatggtcaggtttta 6tgata agacctacgc catgggacat ctaattcaga ggaagaaggt ccatgtcttt 66gaat tgagtctggt gactttgttt cgatgtattc aaaatatgcc tgaaacacta 72aatt cctgctattc agctggcatt gcaaaactgg aagaaggaga tgaactccaa 78atac caagagaaaa tgcacaaatatcactggatg gagatgtcac attttttggt 84aaac tgctgtga 85828285PRThomo sapiens 28Met Asp Asp Ser Thr Glu Arg Glu Gln Ser Arg Leu Thr Ser Cys Leuys Arg Glu Glu Met Lys Leu Lys Glu Cys Val Ser Ile Leu Pro 2Arg Lys Glu Ser Pro Ser ValArg Ser Ser Lys Asp Gly Lys Leu Leu 35 4 Ala Thr Leu Leu Leu Ala Leu Leu Ser Cys Cys Leu Thr Val Val 5Ser Phe Tyr Gln Val Ala Ala Leu Gln Gly Asp Leu Ala Ser Leu Arg65 7Ala Glu Leu Gln Gly His His Ala Glu Lys Leu Pro Ala Gly Ala Gly85 9 Pro Lys Ala Gly Leu Glu Glu Ala Pro Ala Val Thr Ala Gly Leu Ile Phe Glu Pro Pro Ala Pro Gly Glu Gly Asn Ser Ser Gln Asn Arg Asn Lys Arg Ala Val Gln Gly Pro Glu Glu Thr Val Thr Gln Cys Leu Gln LeuIle Ala Asp Ser Glu Thr Pro Thr Ile Gln Lys Gly Ser Tyr Thr Phe Val Pro Trp Leu Leu Ser Phe Lys Arg Gly Ser Leu Glu Glu Lys Glu Asn Lys Ile Leu Val Lys Glu Thr Gly Tyr Phe Ile Tyr Gly Gln Val Leu Tyr Thr AspLys Thr Tyr Ala Met 2is Leu Ile Gln Arg Lys Lys Val His Val Phe Gly Asp Glu Leu 222u Val Thr Leu Phe Arg Cys Ile Gln Asn Met Pro Glu Thr Leu225 234n Asn Ser Cys Tyr Ser Ala Gly Ile Ala Lys Leu Glu Glu Gly 24525p Glu Leu Gln Leu Ala Ile Pro Arg Glu Asn Ala Gln Ile Ser Leu 267y Asp Val Thr Phe Phe Gly Ala Leu Lys Leu Leu 275 28homo sapiens 29atgtggttct tgacaactct gctcctttgg gttccagttg atgggcaagt ggacaccaca 6gtgatcactttgca gcctccatgg gtcagcgtgt tccaagagga aaccgtaacc actgtg aggtgctcca tctgcctggg agcagctcta cacagtggtt tctcaatggc ccactc agacctcgac ccccagctac agaatcacct ctgccagtgt caatgacagt 24taca ggtgccagag aggtctctca gggcgaagtg accccatacagctggaaatc 3aggct ggctactact gcaggtctcc agcagagtct tcacggaagg agaacctctg 36aggt gtcatgcgtg gaaggataag ctggtgtaca atgtgcttta ctatcgaaat 42gcct ttaagttttt ccactggaat tctaacctca ccattctgaa aaccaacata 48aatg gcacctacca ttgctcaggcatgggaaagc atcgctacac atcagcagga 54gtca ctgtgaaaga gctatttcca gctccagtgc tgaatgcatc tgtgacatcc 6cctgg aggggaatct ggtcaccctg agctgtgaaa caaagttgct cttgcagagg 66ttgc agctttactt ctccttctac atgggcagca agaccctgcg aggcaggaac 72tctgaataccaaat actaactgct agaagagaag actctgggtt atactggtgc 78gcca cagaggatgg aaatgtcctt aagcgcagcc ctgagttgga gcttcaagtg 84ctcc agttaccaac tcctgtctgg tttcatgtcc ttttctatct ggcagtggga 9gtttt tagtgaacac tgttctctgg gtgacaatac gtaaagaactgaaaagaaag 96tggg atttagaaat ctctttggat tctggtcatg agaagaaggt aatttccagc caagaag acagacattt agaagaagag ctgaaatgtc aggaacaaaa agaagaacag caggaag gggtgcaccg gaaggagccc cagggggcca cgtagcag 74PRThomo sapiens 3p Phe Leu ThrThr Leu Leu Leu Trp Val Pro Val Asp Gly Glnsp Thr Thr Lys Ala Val Ile Thr Leu Gln Pro Pro Trp Val Ser 2Val Phe Gln Glu Glu Thr Val Thr Leu His Cys Glu Val Leu His Leu 35 4 Gly Ser Ser Ser Thr Gln Trp Phe Leu Asn Gly Thr AlaThr Gln 5Thr Ser Thr Pro Ser Tyr Arg Ile Thr Ser Ala Ser Val Asn Asp Ser65 7Gly Glu Tyr Arg Cys Gln Arg Gly Leu Ser Gly Arg Ser Asp Pro Ile 85 9 Leu Glu Ile His Arg Gly Trp Leu Leu Leu Gln Val Ser Ser Arg Phe Thr GluGly Glu Pro Leu Ala Leu Arg Cys His Ala Trp Lys Lys Leu Val Tyr Asn Val Leu Tyr Tyr Arg Asn Gly Lys Ala Phe Phe Phe His Trp Asn Ser Asn Leu Thr Ile Leu Lys Thr Asn Ile Ser His Asn Gly Thr Tyr His Cys Ser GlyMet Gly Lys His Arg Tyr Ser Ala Gly Ile Ser Val Thr Val Lys Glu Leu Phe Pro Ala Pro Leu Asn Ala Ser Val Thr Ser Pro Leu Leu Glu Gly Asn Leu Val 2eu Ser Cys Glu Thr Lys Leu Leu Leu Gln Arg Pro Gly Leu Gln 222r Phe Ser Phe Tyr Met Gly Ser Lys Thr Leu Arg Gly Arg Asn225 234r Ser Glu Tyr Gln Ile Leu Thr Ala Arg Arg Glu Asp Ser Gly 245 25u Tyr Trp Cys Glu Ala Ala Thr Glu Asp Gly Asn Val Leu Lys Arg 267o Glu Leu GluLeu Gln Val Leu Gly Leu Gln Leu Pro Thr Pro 275 28l Trp Phe His Val Leu Phe Tyr Leu Ala Val Gly Ile Met Phe Leu 29sn Thr Val Leu Trp Val Thr Ile Arg Lys Glu Leu Lys Arg Lys33ys Lys Trp Asp Leu Glu Ile Ser Leu Asp SerGly His Glu Lys Lys 325 33l Ile Ser Ser Leu Gln Glu Asp Arg His Leu Glu Glu Glu Leu Lys 345n Glu Gln Lys Glu Glu Gln Leu Gln Glu Gly Val His Arg Lys 355 36u Pro Gln Gly Ala Thr 37NAhomo sapiens 3ctct gtgtggctcctcggctttga cagagtgcaa gacgatgact tgcaaaatgt 6tgga acgcaacata gagaccatca tcaacacctt ccaccaatac tctgtgaagc gcaccc agacaccctg aaccaggggg aattcaaaga gctggtgcga aaagatctgc ttttct caagaaggag aataagaatg aaaaggtcat agaacacatc atggaggacc24caaa tgcagacaag cagctgagct tcgaggagtt catcatgctg atggcgaggc 3tgggc ctcccacgag aagatgcacg agggtgacga gggccctggc caccaccata 36gcct cggggagggc accccctaag accacagtgg ccaagatcac agtggccacg 42gcca cagtcatggt ggccacggcc acagccactaatcaggaggc caggccaccc 48tacc caaccagggc cccggggcct gttatgtcaa actgtcttgg ctgtggggct 54tggg gccaaataaa gtctcttcct ccaagtcagt gctctg 58632omo sapiens 32Met Thr Cys Lys Met Ser Gln Leu Glu Arg Asn Ile Glu Thr Ile Ilehr PheHis Gln Tyr Ser Val Lys Leu Gly His Pro Asp Thr Leu 2R>

3n Gly Glu Phe Lys Glu Leu Val Arg Lys Asp Leu Gln Asn Phe 35 4 Lys Lys Glu Asn Lys Asn Glu Lys Val Ile Glu His Ile Met Glu 5Asp Leu Asp Thr Asn Ala Asp Lys Gln Leu Ser Phe Glu Glu Phe Ile65 7Met Leu Met Ala Arg LeuThr Trp Ala Ser His Glu Lys Met His Glu 85 9 Asp Glu Gly Pro Gly His His His Lys Pro Gly Leu Gly Glu Gly Pro33homo sapiens 33atattggagc agcaagaggc tgggaagcca tcacttacct tgcactgaga aagaagacaa 6gtat gcacagcttt cctccactgctgctgctgct gttctggggt gtggtgtctc cttccc agcgactcta gaaacacaag agcaagatgt ggacttagtc cagaaatacc aaaata ctacaacctg aagaatgatg ggaggcaagt tgaaaagcgg agaaatagtg 24tggt tgaaaaattg aagcaaatgc aggaattctt tgggctgaaa gtgactggga 3gatgctgaaaccctg aaggtgatga agcagcccag atgtggagtg cctgatgtgg 36ttgt cctcactgag ggaaaccctc gctgggagca aacacatctg aggtacagga 42atta cacgccagat ttgccaagag cagatgtgga ccatgccatt gagaaagcct 48tctg gagtaatgtc acacctctga cattcaccaa ggtctctgagggtcaagcag 54tgat atcttttgtc aggggagatc atcgggacaa ctctcctttt gatggacctg 6aatct tgctcatgct tttcaaccag gcccaggtat tggaggggat gctcattttg 66atga aaggtggacc aacaatttca gagagtacaa cttacatcgt gttgcggctc 72tcgg ccattctctt ggactctcccattctactga tatcggggct ttgatgtacc 78acac cttcagtggt gatgttcagc tagctcagga tgacattgat ggcatccaag 84atgg acgttcccaa aatcctgtcc agcccatcgg cccacaaacc ccaaaagcgt 9agtaa gctaaccttt gatgctataa ctacgattcg gggagaagtg atgttcttta 96gattctacatgcgc acaaatccct tctacccgga agttgagctc aatttcattt ttttctg gccacaactg ccaaatgggc ttgaagctgc ttacgaattt gccgacagag aagtccg gtttttcaaa gggaataagt actgggctgt tcagggacag aatgtgctac gataccc caaggacatc tacagctcct ttggcttccc tagaactgtgaagcatatcg ctgctct ttctgaggaa aacactggaa aaacctactt ctttgttgct aacaaatact ggtatga tgaatataaa cgatctatgg atccaagtta tcccaaaatg atagcacatg ttcctgg aattggccac aaagttgatg cagttttcat gaaagatgga tttttctatt ttcatgg aacaagacaatacaaatttg atcctaaaac gaagagaatt ttgactctcc aagctaa tagctggttc aactgcagga aaaattgaac attactaatt tgaatggaaa catggtg tgagtccaaa gaaggtgttt tcctgaagaa ctgtctattt tctcagtcat taacctc tagagtcact gatacacaga atataatctt atttatacct cagtttgcattttttac tatttagaat gtagcccttt ttgtactgat ataatttagt tccacaaatg ggtacaa aaagtcaagt ttgtggctta tggattcata taggccagag ttgcaaagat ttccaga gtatgcaact ctgacgttga tcccagagag cagcttcagt gacaaacata tttcaag acagaaagag acaggagacatgagtctttg ccggaggaaa agcagctcaa cacatgt gcagtcactg gtgtcaccct agataggcaa gggataactc ttctaacaca taagtgt tttatgtttg gaataaagtc aaccttgttt ctactgtttt 69PRThomo sapiens 34Met His Ser Phe Pro Pro Leu Leu Leu Leu Leu Phe Trp Gly Val Valis Ser Phe Pro Ala Thr Leu Glu Thr Gln Glu Gln Asp Val Asp 2Leu Val Gln Lys Tyr Leu Glu Lys Tyr Tyr Asn Leu Lys Asn Asp Gly 35 4 Gln Val Glu Lys Arg Arg Asn Ser Gly Pro Val Val Glu Lys Leu 5Lys Gln Met Gln Glu Phe Phe GlyLeu Lys Val Thr Gly Lys Pro Asp65 7Ala Glu Thr Leu Lys Val Met Lys Gln Pro Arg Cys Gly Val Pro Asp 85 9 Ala Gln Phe Val Leu Thr Glu Gly Asn Pro Arg Trp Glu Gln Thr Leu Thr Tyr Arg Ile Glu Asn Tyr Thr Pro Asp Leu Pro Arg Ala Val Asp His Ala Ile Glu Lys Ala Phe Gln Leu Trp Ser Asn Val Pro Leu Thr Phe Thr Lys Val Ser Glu Gly Gln Ala Asp Ile Met Ile Ser Phe Val Arg Gly Asp His Arg Asp Asn Ser Pro Phe Asp Gly Gly Gly AsnLeu Ala His Ala Phe Gln Pro Gly Pro Gly Ile Gly Asp Ala His Phe Asp Glu Asp Glu Arg Trp Thr Asn Asn Phe Arg 2yr Asn Leu His Arg Val Ala Ala His Glu Leu Gly His Ser Leu 222u Ser His Ser Thr Asp Ile Gly Ala LeuMet Tyr Pro Ser Tyr225 234e Ser Gly Asp Val Gln Leu Ala Gln Asp Asp Ile Asp Gly Ile 245 25n Ala Ile Tyr Gly Arg Ser Gln Asn Pro Val Gln Pro Ile Gly Pro 267r Pro Lys Ala Cys Asp Ser Lys Leu Thr Phe Asp Ala Ile Thr 27528r Ile Arg Gly Glu Val Met Phe Phe Lys Asp Arg Phe Tyr Met Arg 29sn Pro Phe Tyr Pro Glu Val Glu Leu Asn Phe Ile Ser Val Phe33rp Pro Gln Leu Pro Asn Gly Leu Glu Ala Ala Tyr Glu Phe Ala Asp 325 33g Asp Glu Val ArgPhe Phe Lys Gly Asn Lys Tyr Trp Ala Val Gln 345n Asn Val Leu His Gly Tyr Pro Lys Asp Ile Tyr Ser Ser Phe 355 36y Phe Pro Arg Thr Val Lys His Ile Asp Ala Ala Leu Ser Glu Glu 378r Gly Lys Thr Tyr Phe Phe Val Ala Asn LysTyr Trp Arg Tyr385 39lu Tyr Lys Arg Ser Met Asp Pro Gly Tyr Pro Lys Met Ile Ala 44sp Phe Pro Gly Ile Gly His Lys Val Asp Ala Val Phe Met Lys 423y Phe Phe Tyr Phe Phe His Gly Thr Arg Gln Tyr Lys Phe Asp 435 44o Lys Thr Lys Arg Ile Leu Thr Leu Gln Lys Ala Asn Ser Trp Phe 456s Arg Lys Asn465352995DNAhomo sapiens 35tcttcctcag catttcctgc caaccctttt ctggtttctg tttcttcaac ctctttttgt 6gtcc cttttatggt attttctctt atttctagga aaacattaatgataggtttt cacttc ccctgagtag tctctgttgc aggttgcttt tttgtttgct ctgccctctg tcatac tagaggcctt gctcaaatgt ctggtgacct tggcaatgtg ctaatattta 24aaga acaaaaaaat taattggtaa ccctgaacat gagtggagct tgtcaactgg 3acttt ttgatagtct ggcagagactttttcttcag gaacttccag cgacaatatc 36gctt ttttcttgag ccaatcagat tctctgagaa gttgcttcct atcttctgcc 42gaaa aatcttgtct ggcagcattc tcgcagctag gtgatgacta aagcagggat 48catt cagtatggaa gcttttgttt aattcacctg tttttcacct gatttccatc 54aagttgatctctta cactctttct agagactaga ccttactctg ctgctgaaaa 6aagag agagaattgg taaaagggtt ttgagactct acttcataaa caagctttaa 66tctt atatctacag cctatcttta ccctcatttc cagaagtacc tggtgctgtc 72tgtg ctatttggag tttcaagata taaatcaggt tatttcttggttttctccat 78ctta ggatttagct ttctctgctc tgtaaagcta tccatcttac tttctaagct 84tttt tggtagggtt ttctctttcc ctatttggct ttgtatattg atgtcttttt 9tttta ctgccattgg agagggtttt caggaaggag cagaggctaa tgcatgcgtg 96acta tctttaactg caagttcaattctaattaat atctattcca tttgcatcac tgattca tgatctcaga tgactcacag agattttacc tacagctgtt atctaggctc tatctat ttttccttcc cggatattgt ttagattcca ttggcttaca tcgagtagag atcagcc atacggaacc tgctgtgagc cttcacttgt acagtccacc ttttgatacacatgcct ttgatcaaag aacaggacat aaaaacaaag tcacaatgac attccatagt tttggaa tcagaactcc aaatgtgagt ataatttctt ctaggtttaa atgctctgta tggcact gttacactgc atgtatgcaa agaatgagat gagtcttaat tcatggataa ccaatat atcttgattg ttttatttaactttagcgac tatcttttca atttattttt aacttat tatgttagat ccaaacatta gttgagaaat ttcaaaatgt atgcttgttt aagaaaa tgttttcatt tacttgggaa atgagagtga aaactattat ttgatataac tactgcc atggcttact atgcataggt ctaatttgct gggccaacag ggataagttgaattaac ctttgtaaag gttattggct gatgatataa ctaaagcaaa ttgtctgttt ttgccaa tgttgcttat tcttagagtt gttatgccat ttataccaca aataagaata tatacta acatgttcct ccttgagaaa caagcatttc ctaccaaaaa taaaaaggca aacacta atatagcagg gcagtcttctacctctgaaa taagtttgtc atttagagga gtcttta attatcagaa tatccagaaa tagacaatgt gtgaaaatgg agacccaagc ccttgag tcttaatgaa agcgcagcct ctgagggaag ccagggcaga attttctcat cgctccg cttcatcttt atcttccaca ttccaaaccc taagagatat tcttattgcg2aaacaa caaggctccc ttgcccttgc tgagttttca tacactgcta caaaagcaga 2tttaga aggagttaaa atgtcttttg taaagtttct tacacagctt tctttttttc 2ttttac aggcaacttc gggctcgctg gagaacaact aaggggcacc aaagccctct 222ttac tttaaggttc gctgtatgtttgccttggac aaaaaggcta cctaccacgt 228cagt aatatactta aataagccaa tacttagatc tactgtaagg cagatgctaa 234ggca ttaagtaagc aaatagtgcc ctcagctact gcagaagaaa agtcccactg 24aagaa agtcttgtga tttttaaagg caagttttca agtgctctca tagttctatc246ttcc attaaatcca tactaggagc gtcagtgagg gttttcatag cttttggaaa 252ggtc tctgaactgt aattagcgca agaagtaaaa acagaaacgt caaacgtcaa 258gctt tgttacctgg aggactaaat gtagatgtct ttagtatact ttgtatgttc 264attg gaagataatt ttgtgaatctgtagatttta ttttttcagt cttaccttac 27tcttt tctatgaata atagaggaac tcacggcact ctgccacttg ttaatgaaag 276caga ggatttagaa aagtacatga tccccagacc acaacaaacc aaaacataaa 282tctg tgtcccatgg tcatagtcaa agattttgta ctgctaaaat taccaaataa288taaa gtggatttga acacaatttg aaggtgtctt tctgattaac atgatagaaa 294ataa atcagtttct tagatctaga tatacaaaag cactgtgaca aatgg 2995362mo sapiens 36Met Glu Gln Thr Glu Val Leu Lys Pro Arg Thr Leu Ala Asp Leu Ilele Leu His GlnLeu Phe Ala Gly Asp Glu Val Asn Val Glu Glu 2Val Gln Ala Ile Met Glu Ala Tyr Glu Ser Asp Pro Ile Glu Trp Ala 35 4 Tyr Ala Lys Phe Asp Gln Tyr Arg Tyr Thr Arg Asn Leu Val Asp 5Gln Gly Asn Gly Lys Phe Asn Leu Met Ile Leu Cys Trp GlyGlu Gly65 7His Gly Ser Ser Ile His Asp His Thr Asn Ser His Cys Phe Leu Lys 85 9 Leu Gln Gly Asn Leu Lys Glu Thr Leu Phe Ala Trp Pro Asp Lys Ser Asn Glu Met Val Lys Lys Ser Glu Arg Val Leu Arg Glu Asn Cys AlaTyr Ile Asn Asp Ser Ile Gly Leu His Arg Val Glu Asn Ser His Thr Glu Pro Ala Val Ser Leu His Leu Tyr Ser Pro Pro Phe Asp Thr Cys His Ala Phe Asp Gln Arg Thr Gly His Lys Asn Lys Thr Met Thr Phe His Ser Lys PheGly Ile Arg Thr Pro Asn Ala Ser Gly Ser Leu Glu Asn Asn 37homo sapiens 37gcactggact gagaaccttc accaaaaaaa tgtctgccca gagacagatg aggtccttca 6gtgc tgattggttc ttttccaaag gcccatctaa tcctaccacg cacggaaata caggtttttattcttt ctgccagcta catcagatcc atcaggtccg agctgagttg ccacta cttttccctt tgtctcaatt atgtcttgga agaaggcttt gcggatcccc 24cttc gggcaccaac tgtgaccttg atgctggcga tgctgagcac cccagtggct 3cagag accctcccga ggatttcgtg ctccagttta aggccatgtgctacttcacc 36acgg agcgcgtgcg ttatgtgacc agatacatct ataaccgaga ggaggacgtg 42gaca gcgacgtggg ggtgtatcgg gcggtgacgg cgcaggggcg gcctgacgcc 48tgga acagccagaa ggacatcctg gagaggaccc gagcggagtt ggacacggtg 54caca actacgaggt ggcgttccgcgggatcttgc agaggagagt ggagcccaca 6catct ccccatccag gacagaggcc ctcaaccacc acaacctgct ggtctgctcg 66gatt tctatccagg ccagatcaaa gtccggtggt ttcggaatga ccaggaggag 72ggcg ttgtgtccac cccccttatt aggaacggtg actggacctt ccagatcctg 78ctggaaatgactcc ccagcatgga gacgtctaca cctgccacgt ggagcacccc 84caga gccccatcac cgtggagtgg cgggctcagt ctgaatctgc ccagaacaag 9gagtg gcattggagg cttcgtgctg gggctgatct tcctcgggct gggccttatc 96caaa ggagtcagaa aggacctcaa gggcctccac cagcagggcttctgcactga ctgagac tattttaact aggattggtt atcactcttc tgtgatgcct gcttgtgcct cagaatt cccagctgcc tgtgtcagct tgtcccccga gatcaaagtc ctacagtggc cacgcag ccaccaggtc atctcctttc atccccaccc caaggcgctg gctgtgactc ttcctgc actgacccagagcctctgcc tgtgcacggc cagctgcgtc tactcaggtc aggggtt tctgtttcta ttctctcctc agactgctca agagaagcac atgaaaaaca cctgact ttagagcttt tttacataat taaacatgat cctgagttaa aaaaaaaaaa aatcgct gcagaatgaa ggaatatccc ttgaggtgac ccagccaacc tgtggccagaagggttg taccttgaaa agaccactga aagcattttg gggtgtcaag taagggtggg aggaggt agaaaatcaa ttcaattgtc gcatcattca tggttcttta atattgatgc gtgcatt ggccttagaa tatcccagcc tctcttctgg tttggtgagt gctgtgtaag gcatggt agaattgttt ggagacatatatagtgatcc ttggtcactg gtgtttcaaa tctggaa agtcacatcg atcaagaata ttttttattt ttaagaaagc ataaccagca aaaatac tatttttgag tct 69PRThomo sapiens 38Met Ser Trp Lys Lys Ala Leu Arg Ile Pro Gly Gly Leu Arg Ala Proal Thr Leu MetLeu Ala Met Leu Ser Thr Pro Val Ala Glu Gly 2Arg Asp Pro Pro Glu Asp Phe Val Leu Gln Phe Lys Ala Met Cys Tyr 35 4 Thr Asn Gly Thr Glu Arg Val Arg Tyr Val Thr Arg Tyr Ile Tyr 5Asn Arg Glu Glu Asp Val Arg Phe Asp Ser Asp Val Gly ValTyr Arg65 7Ala Val Thr Ala Gln Gly Arg Pro Asp Ala Glu Tyr Trp Asn Ser Gln 85 9 Asp Ile Leu Glu Arg Thr Arg Ala Glu Leu Asp Thr Val Cys Arg Asn Tyr Glu Val Ala Phe Arg Gly Ile Leu Gln Arg Arg Val Glu Thr ValThr Ile Ser Pro Ser Arg Thr Glu Ala Leu Asn His His Leu Leu Val Cys Ser Val Thr Asp Phe Tyr Pro Gly Gln Ile Lys Val Arg Trp Phe Arg Asn Asp Gln Glu Glu Thr Ala Gly Val Val Ser Pro Leu Ile Arg Asn Gly Asp TrpThr Phe Gln Ile Leu Val Met Glu Met Thr Pro Gln His Gly Asp Val Tyr Thr Cys His Val Glu 2ro Ser Leu Gln Ser Pro Ile Thr Val Glu Trp Arg Ala Gln Ser 222r Ala Gln Asn Lys Met Leu Ser Gly Ile Gly Gly Phe ValLeu225 234u Ile Phe Leu Gly Leu Gly Leu Ile Ile Arg Gln Arg Ser Gln 245 25s Gly Pro Gln Gly Pro Pro Pro Ala Gly Leu Leu His 26homo sapiens 39gaattcggct gtcgcactta ctgttcaata gtatatactc tgtatttgaa aaatagatgt 6tctaggtgataaat taaaaatgaa agaatttaat cattggaaag tattaaatat tgctta tcttctccaa ggaagaggag ttctctcgta cccatccaaa ctgacctaat aagctg cttcatcttg cttgtactgt aggttcattt gcaatttgta gattatgctc 24gatt ggcttttgta aatttctgtt agaagctggt ttctgcatttttgatttttg 3ttgga tacattttca tattgtgcag agaaatccat gagttaaaaa attatttttc 36ttat ttctgcatga acctaagtca cattgaccca gtaattgata tatgtgtgat 42aatt aagtataaga aggtagaata tatagtttta ttagacagat gcttcctgaa 48tttt gtatgttttt actatatcctttttgtgtat ctacagatac aacagacatg 54aatg gactcagaaa tatgcaatgt aaaaatcaaa aacattttca tatataacca 6ctgta aaatctaggt tttttttcaa cattagcagt aaattgagca ctgtttacct 66ttgt accatgaaac catttgattt ttaccatttt aaatgtgtct caagcaagac 72aacttccaaaaata cccttaagac tgtgatgaga gcatttatca ttttgtatgc 78aaag acatttatta tggtttttaa gatacttgga catctgcatc ttcagcttac 84taca atgcagctga aaaagcaacc aaattatttt ttgctgaaaa ctagatgttt 9atgag aaaatactgt atgtgtgtct aagatgtcag ttttataaatctgtattcag 96tcct ttgttagctc actttataat ttgtattttt tttctgtata gaactaaata tctattt acatgtatgt caactcatta cttttttcct gtgaacagta ttgaaaaccc ccggctg ataattaagt gaattaactg tgtctccctt gtcttaggat attctgtaga attgcag atttcttaaatctgaaatga ctttacactg taattctcag catactgatt gagaaca cttgttttga attttgttat acttgactta actttattgc aatgtgaatt tgactgc taagtaggaa gatgtgtaac ttttatttgt tgctattcac atttgaattt cctgtat aggcaatatt atattgacac cttttacaga tcttactgta gcaaaaaccaaaataaa atgctttttc tgct 38PRThomo sapiens 4s Glu Phe Asn His Trp Lys Val Leu Asn Ile Tyr Cys Leu Serro Arg Lys Arg Ser Ser Leu Val Pro Ile Gln Thr Asp Leu Ile 2Leu Lys Leu Leu His Leu Ala Cys Thr Val Gly Ser PheAla Ile Cys 35 4 Leu Cys Ser Phe Arg Ile Gly Phe Cys Lys Phe Leu Leu Glu Ala 5Gly Phe Cys Ile Phe Asp Phe Cys Val Phe Gly Tyr Ile Phe Ile Leu65 7Cys Arg Glu Ile His

Glu Leu Lys Asn Tyr Phe Ser Leu Phe Tyr Phe 85 9 Met Asn Leu Ser His Ile Asp Pro Val Ile Asp Ile Cys Val Ile Ala Ile Lys Tyr Lys Lys Val Glu Tyr Ile Val Leu Leu Asp Arg Phe Leu Lys Tyr Tyr Phe Val Cys Phe TyrTyr Ile Leu Phe Val Leu Gln Ile Gln Gln Thr Cys Lys Arg Met Asp Ser Glu Ile Cys Asn Val Lys Ile Lys Asn Ile Phe Ile Tyr Asn Gln Ser Thr Val Lys Arg Phe Phe Phe Asn Ile Ser Ser Lys Leu Ser Thr Val Tyr Leu His Cys Thr Met Lys Pro Phe Asp Phe Tyr His Phe Lys Cys Val 2er Lys Thr Lys Gln Thr Ser Lys Asn Thr Leu Lys Thr Val Met 222a Phe Ile Ile Leu Tyr Ala Leu Arg Lys Thr Phe Ile Met Val225 234s Ile Leu GlyHis Leu His Leu Gln Leu Thr Arg Ser Thr Met 245 25n Leu Lys Lys Gln Pro Asn Tyr Phe Leu Leu Lys Thr Arg Cys Phe 267s Glu Lys Ile Leu Tyr Val Cys Leu Arg Cys Gln Phe Tyr Lys 275 28r Val Phe Arg Phe His Pro Leu Leu Ala His PheIle Ile Cys Ile 29he Leu Tyr Arg Thr Lys Tyr Ile Leu Phe Thr Cys Met Ser Thr33is Tyr Phe Phe Pro Val Asn Ser Ile Glu Asn Pro Asn Arg Leu Ile 325 33e Lys4Ahomo sapiens 4gtgc tagaagccag tgctcgtgaa ctaaggagaaaaagaacaga caagggaaca 6acat ggcatcagag atccacatga caggcccaat gtgcctcatt gagaacacta gcgact gatggcgaat ccagaagctc tgaagatcct ttctgccatt acacagccta ggtggt ggcaattgtg ggcctctacc gcacaggcaa atcctacctg atgaacaagc 24gaaa gaaaaagggcttctctctgg gctccacggt gcagtctcac actaaaggaa 3atgtg gtgtgtgccc caccccaaga agccaggcca catcctagtt ctgctggaca 36gtct gggagatgta gagaagggtg acaaccagaa tgactcctgg atcttcgccc 42tcct cctgagcagc accttcgtgt acaatagcat aggaaccatc aaccagcagg48acca actgtactat gtgacagagc tgacacatag aatccgatca aaatcctcac 54agaa tgagaatgag gttgaggatt cagctgactt tgtgagcttc ttcccagact 6tggac actgagagat ttctccctgg acttggaagc agatggacaa cccctcacac 66agta cctgacatac tccctgaagc tgaagaaaggtaccagtcaa aaagatgaaa 72acct gcccagactc tgtatccgga aattcttccc aaagaaaaaa tgctttgtct 78ggcc cgttcaccgc aggaagcttg cccagctcga gaaactacaa gatgaagagc 84ccga atttgtgcaa caagtagcag acttctgttc ctacatcttt agtaattcca 9aaaac tctttcaggaggcatccagg tcaacgggcc tcgtctagag agcctggtgc 96acgt caatgccatc agcagtgggg atctgccgtg catggagaac gcagtcctgg tggccca gatagagaac tcagctgcag tgcaaaaggc tattgcccac tatgaacagc tgggcca gaaggtgcag ctgcccacag aaagcctcca ggagctgctg gacctgcacaacagtga gagagaggcc attgaagtct tcatcaggag ttccttcaaa gatgtggacc tatttca aaaggagtta gcggcccagc tagaaaaaaa gcgggatgac ttttgtaaac atcagga agcatcatca gatcgttgct caggtttact tcaggtcatt ttcagtcctc aagaaga agtgaaggcg ggaatttattcgaaaccagg gggctatcgt ctctttgttc agctaca agacctgaag aaaaagtact atgaggaacc gaggaagggg atacaggctg agattct gcagacatac ttgaaatcca aggagtctat gactgatgca attctccaga accagac tctcacagaa aaagaaaagg agattgaagt ggaacgtgtg aaagctgagtcacaggc ttcagcaaaa atgttgcagg aaatgcaaag aaagaatgag cagatgatgg agaagga gaggagttat caggaacact tgaaacaact gactgagaag atggagaacg gggtcca gttgctgaaa gagcaagaga ggaccctcgc tcttaaactt caggaacagg aactact aaaagaggga tttcaaaaagaaagcagaat aatgaaaaat gagatacagg tccagac gaaaatgaga cgacgaaagg catgtaccat aagctaaaga ccagagcctt gtcaccc ctaaccaagg cataattgaa acaattttag aatttggaac aagcgtcact tttgata ataattagat cttgcatcat aacaccaaaa gtttataaag gcatgtggtatgatcaa aatcatgttt tttcttaaaa aaaaaaaaaa gactgtaaat tgtgcaacaa 2gcattt acctctgtat caactcagga aatctcataa gctggtacca ctcaggagaa 2attctt ccagatgacc agcagtagac aaatggatac tgagcagagt cttaggtaaa 2ttggga aatatttggg cattggtctggccaagtcta caatgtccca atatcaagga 222ccct agcttcttag tgaagacaat gtacagttat ccattagatc aagactacac 228tgag caataatgtg atttctggac attgcccatg tataatcctc actgatgatt 234taaa gcaaaccacc ttatacagag atctagaatc tctttatgtt ctccagagga24gaaga aaccatgggc aggagtagga attgagtgat aaacaattgg gctaatgaag 246tctc ttattgttca gttcatccag attataactt caatgggaca ctttagacca 252aatt gacactggat taaacaaatt cacataatgc caaatacaca atgtatttat 258gtat aatttgcaaa gatggactttaaaagatgct gtgtaactaa actgaaataa 264tact tattatttag aatgttaaag cttatgatag tcttttctaa ttcttaacac 27cttga aatctttccg agtttcccca gaagagaata tgggattttt tttgacattt 276catt taataatgct cttgtgttta cctagtatat gtagactttg tcttatgtgt282tcct aggaaagtgg ttgatgtttc ttatagcaat taaaaattat ttttgaactg 28842592PRThomo sapiens 42Met Ala Ser Glu Ile His Met Thr Gly Pro Met Cys Leu Ile Glu Asnsn Gly Arg Leu Met Ala Asn Pro Glu Ala Leu Lys Ile Leu Ser 2Ala IleThr Gln Pro Met Val Val Val Ala Ile Val Gly Leu Tyr Arg 35 4 Gly Lys Ser Tyr Leu Met Asn Lys Leu Ala Gly Lys Lys Lys Gly 5Phe Ser Leu Gly Ser Thr Val Gln Ser His Thr Lys Gly Ile Trp Met65 7Trp Cys Val Pro His Pro Lys Lys Pro Gly HisIle Leu Val Leu Leu 85 9 Thr Glu Gly Leu Gly Asp Val Glu Lys Gly Asp Asn Gln Asn Asp Trp Ile Phe Ala Leu Ala Val Leu Leu Ser Ser Thr Phe Val Tyr Ser Ile Gly Thr Ile Asn Gln Gln Ala Met Asp Gln Leu Tyr Tyr Thr Glu Leu Thr His Arg Ile Arg Ser Lys Ser Ser Pro Asp Glu Asn Glu Asn Glu Val Glu Asp Ser Ala Asp Phe Val Ser Phe Phe Pro Phe Val Trp Thr Leu Arg Asp Phe Ser Leu Asp Leu Glu Ala Asp Gln Pro Leu Thr ProAsp Glu Tyr Leu Thr Tyr Ser Leu Lys Leu 2ys Gly Thr Ser Gln Lys Asp Glu Thr Phe Asn Leu Pro Arg Leu 222e Arg Lys Phe Phe Pro Lys Lys Lys Cys Phe Val Phe Asp Arg225 234l His Arg Arg Lys Leu Ala Gln Leu Glu LysLeu Gln Asp Glu 245 25u Leu Asp Pro Glu Phe Val Gln Gln Val Ala Asp Phe Cys Ser Tyr 267e Ser Asn Ser Lys Thr Lys Thr Leu Ser Gly Gly Ile Gln Val 275 28n Gly Pro Arg Leu Glu Ser Leu Val Leu Thr Tyr Val Asn Ala Ile 29er Gly Asp Leu Pro Cys Met Glu Asn Ala Val Leu Ala Leu Ala33ln Ile Glu Asn Ser Ala Ala Val Gln Lys Ala Ile Ala His Tyr Glu 325 33n Gln Met Gly Gln Lys Val Gln Leu Pro Thr Glu Ser Leu Gln Glu 345u Asp Leu His ArgAsp Ser Glu Arg Glu Ala Ile Glu Val Phe 355 36e Arg Ser Ser Phe Lys Asp Val Asp His Leu Phe Gln Lys Glu Leu 378a Gln Leu Glu Lys Lys Arg Asp Asp Phe Cys Lys Gln Asn Gln385 39la Ser Ser Asp Arg Cys Ser Gly Leu Leu GlnVal Ile Phe Ser 44eu Glu Glu Glu Val Lys Ala Gly Ile Tyr Ser Lys Pro Gly Gly 423g Leu Phe Val Gln Lys Leu Gln Asp Leu Lys Lys Lys Tyr Tyr 435 44u Glu Pro Arg Lys Gly Ile Gln Ala Glu Glu Ile Leu Gln Thr Tyr 456s Ser Lys Glu Ser Met Thr Asp Ala Ile Leu Gln Thr Asp Gln465 478u Thr Glu Lys Glu Lys Glu Ile Glu Val Glu Arg Val Lys Ala 485 49u Ser Ala Gln Ala Ser Ala Lys Met Leu Gln Glu Met Gln Arg Lys 55lu Gln Met Met GluGln Lys Glu Arg Ser Tyr Gln Glu His Leu 5525Lys Gln Leu Thr Glu Lys Met Glu Asn Asp Arg Val Gln Leu Leu Lys 534n Glu Arg Thr Leu Ala Leu Lys Leu Gln Glu Gln Glu Gln Leu545 556s Glu Gly Phe Gln Lys Glu Ser Arg Ile MetLys Asn Glu Ile 565 57n Asp Leu Gln Thr Lys Met Arg Arg Arg Lys Ala Cys Thr Ile Ser 589DNAhomo sapiens 43attaaacctc tcgccgagcc cctccgcaga ctctgcgccg gaaagtttca tttgctgtat 6ctcg agagctgtct aggttaacgt tcgcactctg tgtatataacctcgacagtc caccta acgtgctgtg cgtagctgct cctttggttg aatccccagg cccttgttgg caaggt ggcaggatgt ctcagtggta cgaacttcag cagcttgact caaaattcct 24ggtt caccagcttt atgatgacag ttttcccatg gaaatcagac agtacctggc 3ggtta gaaaagcaag actgggagcacgctgccaat gatgtttcat ttgccaccat 36tcat gacctcctgt cacagctgga tgatcaatat agtcgctttt ctttggagaa 42cttg ctacagcata acataaggaa aagcaagcgt aatcttcagg ataattttca 48ccca atccagatgt ctatgatcat ttacagctgt ctgaaggaag aaaggaaaat 54aaacgcccagagat ttaatcaggc tcagtcgggg aatattcaga gcacagtgat 6acaaa cagaaagagc ttgacagtaa agtcagaaat gtgaaggaca aggttatgtg 66gcat gaaatcaaga gcctggaaga tttacaagat gaatatgact tcaaatgcaa 72gcag aacagagaac acgagaccaa tggtgtggca aagagtgatcagaaacaaga 78gtta ctcaagaaga tgtatttaat gcttgacaat aagagaaagg aagtagttca 84aata gagttgctga atgtcactga acttacccag aatgccctga ttaatgatga 9tggag tggaagcgga gacagcagag cgcctgtatt ggggggccgc ccaatgcttg 96tcag ctgcagaact ggttcactatagttgcggag agtctgcagc aagttcggca gcttaaa aagttggagg aattggaaca gaaatacacc tacgaacatg accctatcac aaacaaa caagtgttat gggaccgcac cttcagtctt ttccagcagc tcattcagag gtttgtg gtggaaagac agccctgcat gccaacgcac cctcagaggc cgctggtcttgacaggg gtccagttca ctgtgaagtt gagactgttg gtgaaattgc aagagctgaa taatttg aaagtcaaag tcttatttga taaagatgtg aatgagagaa atacagtaaa atttagg aagttcaaca ttttgggcac gcacacaaaa gtgatgaaca tggaggagtc caatggc agtctggcgg ctgaatttcggcacctgcaa ttgaaagaac agaaaaatgc caccaga acgaatgagg gtcctctcat cgttactgaa gagcttcact cccttagttt aacccaa ttgtgccagc ctggtttggt aattgacctc gagacgacct ctctgcccgt ggtgatc tccaacgtca gccagctccc gagcggttgg gcctccatcc tttggtacaagctggtg gcggaaccca ggaatctgtc cttcttcctg actccaccat gtgcacgatg tcagctt tcagaagtgc tgagttggca gttttcttct gtcaccaaaa gaggtctcaa ggaccag ctgaacatgt tgggagagaa gcttcttggt cctaacgcca gccccgatgg cattccg tggacgaggt tttgtaaggaaaatataaat gataaaaatt ttcccttctg ttggatt gaaagcatcc tagaactcat taaaaaacac ctgctccctc tctggaatga gtgcatc atgggcttca tcagcaagga gcgagagcgt gccctgttga aggaccagca ggggacc ttcctgctgc ggttcagtga gagctcccgg gaaggggcca tcacattcac2gtggag cggtcccaga acggaggcga acctgacttc catgcggttg aaccctacac 2aaagaa ctttctgctg ttactttccc tgacatcatt cgcaattaca aagtcatggc 2gagaat attcctgaga atcccctgaa gtatctgtat ccaaatattg acaaagacca 222tgga aagtattact ccaggccaaaggaagcacca gagccaatgg aacttgatgg 228agga actggatata tcaagactga gttgatttct gtgtctgaag ttcacccttc 234tcag accacagaca acctgctccc catgtctcct gaggagtttg acgaggtgtc 24tagtg ggctctgtag aattcgacag tatgatgaac acagtataga gcatgaattt246tctt ctctggcgac agttttcctt ctcatctgtg attccctcct gctactctgt 252acat cctgtgtttc tagggaaatg aaagaaaggc cagcaaattc gctgcaacct 258agca agtgaatttt tctctaactc agaaacatca gttactctga agggcatcat 264tact gaaggtaaaa ttgaaaggcattctctgaag agtgggtttc acaagtgaaa 27ccaga tacacccaaa gtatcaggac gagaatgagg gtcctttggg aaaggagaag 276aaca tctagcaaat gttatgcata aagtcagtgc ccaactgtta taggttgttg 282tcag tggttattta gggaactgct tgacgtagga acggtaaatt tctgtgggag288taca tgttttcttt gctttaagtg taactggcag ttttccattg gtttacctgt 294gttc aaagccaagt ttatatacaa ttatatcagt cctctttcaa aggtagccat 3gatctg gtagggggaa aatgtgtatt ttattacatc tttcacattg gctatttaaa 3aagaca aattctgttt cttgagaagagaatattagc tttactgttt gttatggctt 3acacta gctaatatca atagaaggat gtacatttcc aaattcacaa gttgtgtttg 3ccaaag ctgaatacat tctgctttca tcttggtcac atacaattat ttttacagtt 324aggg agttaggcta ttcacaacca ctcattcaaa agttgaaatt aaccatagat33taaac tcagaaattt aattcatgtt tcttaaatgg gctactttgt cctttttgtt 336gtgg tatttagtct attagccaca aaattgggaa aggagtagaa aaagcagtaa 342actt gaataataca ccagagataa tatgagaatc agatcatttc aaaactcatt 348gtaa ctgcattgag aactgcatatgtttcgctga tatatgtgtt tttcacattt 354ggtt ccattctctc tcctgtactt tttccagaca cttttttgag tggatgatgt 36gaagt atactgtatt tttacctttt tccttcctta tcactgacac aaaaagtaga 366gatg ggtttgacaa ggttcttccc ttttacatac tgctgtctat gtggctgtat372tttc cactactgct accacaacta tattatcatg caaatgctgt attcttcttt 378gata aagatttctt gagttttgtt ttaaaattaa agctaaagta tctgtattgc 384tata atatcgacac agtgctttcc gtggcactgc atacaatctg aggcctcctc 39gtttt tatatagatg gcgagaacctaagtttcagt tgattttaca attgaaatga 396aaca aagaagacaa cattaaaaac aatattgttt cta 4o sapiens 44Met Ser Gln Trp Tyr Glu Leu Gln Gln Leu Asp Ser Lys Phe Leu Glual His Gln Leu Tyr Asp Asp Ser Phe Pro Met Glu Ile Arg Gln 2Tyr Leu Ala Gln Trp Leu Glu Lys Gln Asp Trp Glu His Ala Ala Asn 35 4 Val Ser Phe Ala Thr Ile Arg Phe His Asp Leu Leu Ser Gln Leu 5Asp Asp Gln Tyr Ser Arg Phe Ser Leu Glu Asn Asn Phe Leu Leu Gln65 7His Asn Ile Arg Lys Ser Lys ArgAsn Leu Gln Asp Asn Phe Gln Glu 85 9 Pro Ile Gln Met Ser Met Ile Ile Tyr Ser Cys Leu Lys Glu Glu Lys Ile Leu Glu Asn Ala Gln Arg Phe Asn Gln Ala Gln Ser Gly Ile Gln Ser Thr Val Met Leu Asp Lys Gln Lys Glu Leu Asp Ser Val Arg Asn Val Lys Asp Lys Val Met Cys Ile Glu His Glu Ile Lys Ser Leu Glu Asp Leu Gln Asp Glu Tyr Asp Phe Lys Cys Lys Thr Gln Asn Arg Glu His Glu Thr Asn Gly Val Ala Lys Ser Asp Gln Gln Glu GlnLeu Leu Leu Lys Lys Met Tyr Leu Met Leu Asp Asn 2rg Lys Glu Val Val His Lys Ile Ile Glu Leu Leu Asn Val Thr 222u Thr Gln Asn Ala Leu Ile Asn Asp Glu Leu Val Glu Trp Lys225 234g Gln Gln Ser Ala Cys Ile Gly GlyPro Pro Asn Ala Cys Leu 245 25p Gln Leu Gln Asn Trp Phe Thr Ile Val Ala Glu Ser Leu Gln Gln 267g Gln Gln Leu Lys Lys Leu Glu Glu Leu Glu Gln Lys Tyr Thr 275 28r Glu His Asp Pro Ile Thr Lys Asn Lys Gln Val Leu Trp Asp Arg 29he Ser Leu Phe Gln Gln Leu Ile Gln Ser Ser Phe Val Val Glu33rg Gln Pro Cys Met Pro Thr His Pro Gln Arg Pro Leu Val Leu Lys 325 33r Gly Val Gln Phe Thr Val Lys Leu Arg Leu Leu Val Lys Leu Gln 345u Asn Tyr AsnLeu Lys Val Lys Val Leu Phe Asp Lys Asp Val 355 36n Glu Arg Asn Thr Val Lys Gly Phe Arg Lys Phe Asn Ile Leu Gly 378s Thr Lys Val Met Asn Met Glu Glu Ser Thr Asn Gly Ser Leu385 39la Glu Phe Arg His Leu Gln Leu Lys GluGln Lys Asn Ala Gly 44rg Thr Asn Glu Gly Pro Leu Ile Val Thr Glu Glu Leu His Ser 423r Phe Glu Thr Gln Leu Cys Gln Pro Gly Leu Val Ile Asp Leu 435 44u Thr Thr Ser Leu Pro Val Val Val Ile Ser Asn Val Ser Gln Leu 456r Gly Trp Ala Ser Ile Leu Trp Tyr Asn Met

Leu Val Ala Glu465 478g Asn Leu Ser Phe Phe Leu Thr Pro Pro Cys Ala Arg Trp Ala 485 49n Leu Ser Glu Val Leu Ser Trp Gln Phe Ser Ser Val Thr Lys Arg 55eu Asn Val Asp Gln Leu Asn Met Leu Gly Glu Lys Leu Leu Gly5525Pro Asn Ala Ser Pro Asp Gly Leu Ile Pro Trp Thr Arg Phe Cys Lys 534n Ile Asn Asp Lys Asn Phe Pro Phe Trp Leu Trp Ile Glu Ser545 556u Glu Leu Ile Lys Lys His Leu Leu Pro Leu Trp Asn Asp Gly 565 57s Ile Met GlyPhe Ile Ser Lys Glu Arg Glu Arg Ala Leu Leu Lys 589n Gln Pro Gly Thr Phe Leu Leu Arg Phe Ser Glu Ser Ser Arg 595 6lu Gly Ala Ile Thr Phe Thr Trp Val Glu Arg Ser Gln Asn Gly Gly 662o Asp Phe His Ala Val Glu Pro Tyr ThrLys Lys Glu Leu Ser625 634l Thr Phe Pro Asp Ile Ile Arg Asn Tyr Lys Val Met Ala Ala 645 65u Asn Ile Pro Glu Asn Pro Leu Lys Tyr Leu Tyr Pro Asn Ile Asp 667p His Ala Phe Gly Lys Tyr Tyr Ser Arg Pro Lys Glu Ala Pro 67568u Pro Met Glu Leu Asp Gly Pro Lys Gly Thr Gly Tyr Ile Lys Thr 69eu Ile Ser Val Ser Glu Val His Pro Ser Arg Leu Gln Thr Thr77sp Asn Leu Leu Pro Met Ser Pro Glu Glu Phe Asp Glu Val Ser Arg 725 73e Val Gly Ser ValGlu Phe Asp Ser Met Met Asn Thr Val 745DNAhomo sapiens 45cccggcgtcc cgtcgagccc agccccgccg ggggcgctcc tcgccgcccg cacgccctcc 6atgt cgtccatcct gcctttcact cccccgatcg tgaagcgcct gctgggctgg agggcg agcagaacgg gcaggaggag aaatggtgcgagaaggcggt caagagcctg agaaac tcaagaagac ggggcagctg gacgagctgg agaaggccat caccacgcag 24aaca ccaagtgcat caccatcccc aggtccctgg atggccggtt gcaggtgtcc 3gaagg ggctccctca tgtcatctac tgccgcctgt ggcgatggcc agacctgcac 36cacg agctgcgggccatggagctg tgtgagttcg ccttcaatat gaagaaggac 42tgcg tgaatcccta ccactaccag agagtagaga caccagttct acctcctgtg 48ccac gccacacaga gatcccggcc gagttccccc cactggacga ctacagccat 54cccg aaaacactaa cttccccgca ggcatcgagc cccagagcaa tattccagag6acccc ctggctacct gagtgaagat ggagaaacca gtgaccacca gatgaaccac 66gacg caggttctcc aaacctatcc ccgaatccga tgtccccagc acataataac 72ctgc agccagttac ctactgcgag ccggccttct ggtgctccat ctcctactac 78aacc agcgcgtcgg ggagacattc cacgcctcgcagccatccat gactgtggat 84accg acccctccaa ttcggagcgc ttctgcctag ggctgctctc caatgtcaac 9tgcag cagtggagct gacacggaga cacatcggaa gaggcgtgcg gctctactac 96gggg aggtcttcgc agagtgcctc agtgacagcg ctatttttgt ccagtctccc tgtaacc agcgctatggctggcacccg gccaccgtct gcaagatccc accaggatgc ctgaaga tcttcaacaa ccaggagttc gctgccctcc tggcccagtc ggtcaaccag tttgagg ctgtctacca gttgacccga atgtgcacca tccgcatgag cttcgtcaaa tggggag cggagtacag gagacagact gtgaccagta ccccctgctg gattgagctgctgaatg ggcctttgca gtggcttgac aaggtcctca cccagatggg ctccccaagc cgctgtt ccagtgtgtc ttagagacat caagtatggt aggggagggc aggcttgggg atggcca tacaggaggt ggagaaaatt ggaactctac tcaacccatt gttgtcaagg aagaaat ctttctccct caactgaaggggtgcaccca cctgttttct gaaacacacg aaaccca gaggtggatg ttatgaacag ctgtgtctgc caaacacatt taccctttgg cactttg aagggcaaga aatggcgtct gctctggtgg cttaagtgag cagaacaggt attacac caccggcacc ctccccccag actctttttt tgagtgacag ctttctgggacacagtc caaccagaaa cgcccctctg tctaggactg cagtgtggag ttcaccttgg ggcgttc taggtaggaa gagcccgcac gatgcagacc tcatgcccag ctctctgacg gtgacag tgcctcttcc agtgaacatt cccagcccag ccccgccccg ttgtgagctg agacttg ggatggggag ggagggagttttgtctgtct ccctcccctc tcagaacata attggga ggtgcgtgtt cagcagaacc tgcacacagg acagcgggaa aaatcgatga ccacctc tttaaaaact cacttacgtt gtcctttttc actttgaaaa gttggaagga 2tgaggc ccagtgcata tgcaatgtat agtgtctatt atcacattaa tctcaaagag2gaatga cggtaagtgt tctcatgaag caggaggccc ttgtcgtggg atggcatttg 2caggca gcaccacact gggtgcgtct ccagtcatct gtaagagctt gctccagatt 222cata cggctatatt ggtttatgta gtcagttgca ttcattaaat caactttatc 228tcaa aaaaaaaaaa aag23PRThomo sapiens 46Met Ser Ser Ile Leu Pro Phe Thr Pro Pro Ile Val Lys Arg Leu Leurp Lys Lys Gly Glu Gln Asn Gly Gln Glu Glu Lys Trp Cys Glu 2Lys Ala Val Lys Ser Leu Val Lys Lys Leu Lys Lys Thr Gly Gln Leu 35 4 Glu LeuGlu Lys Ala Ile Thr Thr Gln Asn Val Asn Thr Lys Cys 5Ile Thr Ile Pro Arg Ser Leu Asp Gly Arg Leu Gln Val Ser His Arg65 7Lys Gly Leu Pro His Val Ile Tyr Cys Arg Leu Trp Arg Trp Pro Asp 85 9 His Ser His His Glu Leu Arg Ala Met Glu LeuCys Glu Phe Ala Asn Met Lys Lys Asp Glu Val Cys Val Asn Pro Tyr His Tyr Gln Val Glu Thr Pro Val Leu Pro Pro Val Leu Val Pro Arg His Thr Ile Pro Ala Glu Phe Pro Pro Leu Asp Asp Tyr Ser His Ser Ile Pro Glu Asn Thr Asn Phe Pro Ala Gly Ile Glu Pro Gln Ser Asn Ile Glu Thr Pro Pro Pro Gly Tyr Leu Ser Glu Asp Gly Glu Thr Ser His Gln Met Asn His Ser Met Asp Ala Gly Ser Pro Asn Leu Ser 2sn Pro Met Ser ProAla His Asn Asn Leu Asp Leu Gln Pro Val 222r Cys Glu Pro Ala Phe Trp Cys Ser Ile Ser Tyr Tyr Glu Leu225 234n Arg Val Gly Glu Thr Phe His Ala Ser Gln Pro Ser Met Thr 245 25l Asp Gly Phe Thr Asp Pro Ser Asn Ser Glu ArgPhe Cys Leu Gly 267u Ser Asn Val Asn Arg Asn Ala Ala Val Glu Leu Thr Arg Arg 275 28s Ile Gly Arg Gly Val Arg Leu Tyr Tyr Ile Gly Gly Glu Val Phe 29lu Cys Leu Ser Asp Ser Ala Ile Phe Val Gln Ser Pro Asn Cys33sn Gln Arg Tyr Gly Trp His Pro Ala Thr Val Cys Lys Ile Pro Pro 325 33y Cys Asn Leu Lys Ile Phe Asn Asn Gln Glu Phe Ala Ala Leu Leu 345n Ser Val Asn Gln Gly Phe Glu Ala Val Tyr Gln Leu Thr Arg 355 36t Cys Thr Ile Arg MetSer Phe Val Lys Gly Trp Gly Ala Glu Tyr 378g Gln Thr Val Thr Ser Thr Pro Cys Trp Ile Glu Leu His Leu385 39ly Pro Leu Gln Trp Leu Asp Lys Val Leu Thr Gln Met Gly Ser 44er Ile Arg Cys Ser Ser Val Ser 4242mo sapiens 47tctttaagat ttgtagctac taagaaagaa aggagctttt tttccttggg ccttcaaact 6accg catgagcctg acggcgcatg gtcttaacat caggctgtgc aggaagaagc tgcaga tggatgccag cacacacaag gaagcagagc tctggcaaca ttgagtcaaa ggacac aacatcagagggacggcaga gaatccttgt gtgtagtctt tggtggcagt 24attg caaggaggga ctttaagact acttctgatt tgcaaagatg gtctgtgctc 3aggct aaagtgactg gacgagacgc actgttggag aaataaaaat gacttcccat 36attg ccatctttgc cctgatgagc ttctgtttag ccactgcagg tccagagcct42ctgt gtgaactgtc acctgtcagt gcctcccatc ctgtccaggc cttgatggag 48actg ttttgtcagg ctgtgccagc agaggcacaa ctgggctgcc acaggaggtg 54ctga atctcgcact gcgccagggg cctggccagc tacagagaga ggtcacactt 6gaatc ccatctcctc agtccacatc caccacaagtctgttgtgtt cctgctcaac 66cacc ccctggtgtg gcatctgaag acagagagac ttgccactgg ggtctccaga 72ttgg tgtctgaggg ttctgtggtc cagttttcat cagcaaactt ctccttgaca 78acag aagaaaggaa cttcccccat ggaaatgaac atctgttaaa ttgggcccga 84tatg gagcagttacttcattcacc gaactcaaga tagcaagaaa catttatatt 9ggggg aagatcaagt gttccctcca aagtgcaaca tagggaagaa ttttctctca 96tacc ttgctgagta ccttcaaccc aaagcagcag aagggtgtgt gatgtccagc ccccaga atgaggaagt acacatcatc gagctaatca cccccaactc taacccctacgctttcc aggtggatat aacaattgat ataagacctt ctcaagagga tcttgaagtg aaaaatc tcatcctgat cttgaagtgc aaaaagtctg tcaactgggt gatcaaatct gatgtta agggaagcct gaaaattatt gctcctaaca gtattggctt tggaaaagag gaaagat ctatgacaat gaccaaatcaataagagatg acattccttc aacccaaggg ctggtga agtgggcttt ggacaatggc tatagtccaa taacttcata cacaatggct gtggcaa tagtatttca tcttcggctt gaaaataatg aggagatggg agatgaggaa cacacta ttcctcctga gctacggatc ctgctggacc ctggtgccct gcctgccctgaacccgc ccatccgggg aggggaaggc caaaatggag gccttccgtt tcctttccca atttcca ggagagtctg gaatgaagag ggagaagatg ggctccctcg gccaaaggac gtcattc ccagcataca actgtttcct ggtctcagag agccagaaga ggtgcaaggg gtggata ttgccctgtc tgtcaaatgtgacaatgaga agatgatcgt ggctgtagaa gattctt ttcaggccag tggctactcg gggatggacg tcaccctgtt ggatcctacc aaggcca agatgaatgg cacacacttt gttttggagt ctcctctgaa tggctgcggt cggcccc ggtggtcagc ccttgatggt gtggtctact ataactccat tgtgatacagccagccc ttggggacag tagtggttgg ccagatggtt atgaagatct ggagtcaggt aatggat ttccgggaga tatggatgaa ggagatgctt ccctgttcac ccgacctgaa 2tggtgt ttaattgcag ccttcagcag gtgaggaacc ccagcagctt ccaggaacag 2acggaa acatcacctt caacatggagctatacaaca ctgacctctt tttggtgccc 2agggcg tcttctctgt gccagagaat ggacacgttt atgttgaggt atctgttact 222gaac aagaactggg atttgccatc caaacgtgct ttatctctcc atattcgaac 228agga tgtctcatta caccattatt gagaatattt gtcctaaaga tgaatctgtg234taca gtcccaagag agtgcacttc cctatcccgc aagctgacat ggataagaag 24cagct ttgtcttcaa gcctgtcttc aacacctcac tgctctttct acagtgtgag 246ctgt gtacgaagat ggagaagcac ccccagaagt tgcctaagtg tgtgcctcct 252gcct gcacctcgct ggacgcctcgataatctggg ccatgatgca gaataagaag 258acca agccccttgc tgtgatccac catgaagcag aatctaaaga aaaaggtcca 264aagg aaccaaatcc aatttctcca ccaattttcc atggtctgga caccctaacc 27gggca ttgcgtttgc agcctttgtg atcggagcac tcctgacggg ggccttgtgg276tatt ctcacacagg ggagacagca ggaaggcagc aagtccccac ctccccgcca 282gaaa acagcagtgc tgcccacagc atcggcagca cgcagagcac gccttgctcc 288agca cggcctagcc caacccaggc ccaacccggc ccaacccagc ccagcccagc 294cagc tactccaagg gcaggaccaatggctgagcc tcgtgtccag actcagaggg 3attttg gttcccttgt aaagacagag tgaatttcag tataaagatc acccgttgta 3ccccac acccagggct agtataaaca tgaccctggg cttctgtacc acactagaat 3gtgaga aagctaaaat ggtggtcttc tccaccagcc cctcacaggc ttgggggttt3tgtgaa acacatgcca gtttttaaaa tgctgctttg tccaggtgag aacatccata 324ggcc ctgagtttta cccagactca aggagttggt aaagggttaa tagccagata 33accag tgaggagatg cggccaaaga ttctttatat ctgaaccaag atgtaaaaca 336gctt tgaggctttc taagcgatcctcctgtctaa tttgcacctt tgtctggatg 342tctg accttgctgc cacaacctgt ggggtctgat gtgtcccaag atgggtgctg 348ggga ctgcaccctg acaagtgtta aggcaacatt ccttgcttgt gccctgggcc 354aatg ctgatgacct tatcagcttc ctgtttcttc ccatactgca tacaccactg36tgtct taatgcaaat tttgtatttc ttacaggcct acagaaattg aaaatgacca 366ggaa ccacagattt gtgcccattc ctaatatttt gttctgcaaa ttaatgtata 372ggtg aaattcagtt ataaagtcaa ggacgaattt gcacagtgat atatttctat 378gcaa gtacaagtat ataatatgtcacctggcaca ttcattttct cagttgaaga 384aatt tgaaaatgtc cttatgcttt tagagttgca acttaagtat atttggtagg 39tgttt ccactcaaaa tatgtcaact taaaaaaaaa taggcccttt cataaaaacc 396tagc aagatgcaaa tgcatggcaa atctgtcggt ctccagttgg ttatctgaat4tcacca attccaccaa gacagtgctg agattggaaa gggcactcat ttggattgcc 4ttctct tgccttaaat atatcccata tatttaatat gtcaaaaagg gcttgaggtg 4tcatta aatggaataa tatgatgcca ctttgcagct aaaataagct cagtgatacc 42gtt 42PRThomo sapiens 48MetThr Ser His Tyr Val Ile Ala Ile Phe Ala Leu Met Ser Phe Cysla Thr Ala Gly Pro Glu Pro Gly Ala Leu Cys Glu Leu Ser Pro 2Val Ser Ala Ser His Pro Val Gln Ala Leu Met Glu Ser Phe Thr Val 35 4 Ser Gly Cys Ala Ser Arg Gly Thr ThrGly Leu Pro Gln Glu Val 5His Val Leu Asn Leu Ala Leu Arg Gln Gly Pro Gly Gln Leu Gln Arg65 7Glu Val Thr Leu His Leu Asn Pro Ile Ser Ser Val His Ile His His 85 9 Ser Val Val Phe Leu Leu Asn Ser Pro His Pro Leu Val Trp His Lys Thr Glu Arg Leu Ala Thr Gly Val Ser Arg Leu Phe Leu Val Glu Gly Ser Val Val Gln Phe Ser Ser Ala Asn Phe Ser Leu Thr Glu Thr Glu Glu Arg Asn Phe Pro His Gly Asn Glu His Leu Leu Asn Trp Ala Arg Lys GluTyr Gly Ala Val Thr Ser Phe Thr Glu Leu Ile Ala Arg Asn Ile Tyr Ile Lys Val Gly Glu Asp Gln Val Phe Pro Lys Cys Asn Ile Gly Lys Asn Phe Leu Ser Leu Asn Tyr Leu 2lu Tyr Leu Gln Pro Lys Ala Ala Glu Gly Cys ValMet Ser Ser 222o Gln Asn Glu Glu Val His Ile Ile Glu Leu Ile Thr Pro Asn225 234n Pro Tyr Ser Ala Phe Gln Val Asp Ile Thr Ile Asp Ile Arg 245 25o Ser Gln Glu Asp Leu Glu Val Val Lys Asn Leu Ile Leu Ile Leu 267s Lys Lys Ser Val Asn Trp Val Ile Lys Ser Phe Asp Val Lys 275 28y Ser Leu Lys Ile Ile Ala Pro Asn Ser Ile Gly Phe Gly Lys Glu 29lu Arg Ser Met Thr Met Thr Lys Ser Ile Arg Asp Asp Ile Pro33er Thr Gln Gly Asn Leu ValLys Trp Ala Leu Asp Asn Gly Tyr Ser 325 33o Ile Thr Ser Tyr Thr Met Ala Pro Val Ala Ile Val Phe His Leu 345u Glu Asn Asn Glu Glu Met Gly Asp Glu Glu Val His Thr Ile 355 36o Pro Glu Leu Arg Ile Leu Leu Asp Pro Gly Ala Leu ProAla Leu 378n Pro Pro Ile Arg Gly Gly Glu Gly Gln Asn Gly Gly Leu Pro385 39ro Phe Pro Asp Ile Ser Arg Arg Val Trp Asn Glu Glu Gly Glu 44ly Leu Pro Arg Pro Lys Asp Pro Val Ile Pro Ser Ile Gln Leu 423oGly Leu Arg Glu Pro Glu Glu Val Gln Gly Ser Val Asp Ile 435 44a Leu Ser Val Lys Cys Asp Asn Glu Lys Met Ile Val Ala Val Glu 456p Ser Phe Gln Ala Ser Gly Tyr Ser Gly Met Asp Val Thr Leu465 478p Pro Thr Cys Lys Ala LysMet Asn Gly Thr His Phe Val Leu 485 49u Ser Pro Leu Asn Gly Cys Gly Thr Arg Pro Arg Trp Ser Ala Leu 55ly Val Val Tyr Tyr Asn Ser Ile Val Ile Gln Val Pro Ala Leu 5525Gly Asp Ser Ser Gly Trp Pro Asp Gly Tyr Glu Asp Leu Glu SerGly 534n Gly Phe Pro Gly Asp Met Asp Glu Gly Asp Ala Ser Leu Phe545 556g Pro Glu Ile Val Val Phe Asn Cys Ser Leu Gln Gln Val Arg 565 57n Pro Ser Ser Phe Gln Glu Gln Pro His Gly Asn Ile Thr Phe Asn 589u LeuTyr Asn Thr Asp Leu Phe Leu Val Pro Ser Gln Gly Val 595 6he Ser Val Pro Glu Asn Gly His Val Tyr Val Glu Val Ser Val Thr 662a Glu Gln Glu Leu Gly Phe Ala Ile Gln Thr Cys Phe Ile Ser625 634r Ser Asn Pro Asp Arg Met SerHis Tyr Thr Ile Ile Glu Asn 645 65e Cys Pro Lys Asp Glu Ser Val Lys Phe Tyr Ser Pro Lys Arg Val 66BR>

67e Pro Ile Pro Gln Ala Asp Met Asp Lys Lys Arg Phe Ser Phe 675 68l Phe Lys Pro Val Phe Asn Thr Ser Leu Leu Phe Leu Gln Cys Glu 69hr Leu Cys Thr Lys Met Glu Lys His Pro Gln Lys Leu Pro Lys77ys Val ProPro Asp Glu Ala Cys Thr Ser Leu Asp Ala Ser Ile Ile 725 73p Ala Met Met Gln Asn Lys Lys Thr Phe Thr Lys Pro Leu Ala Val 745s His Glu Ala Glu Ser Lys Glu Lys Gly Pro Ser Met Lys Glu 755 76o Asn Pro Ile Ser Pro Pro Ile Phe HisGly Leu Asp Thr Leu Thr 778t Gly Ile Ala Phe Ala Ala Phe Val Ile Gly Ala Leu Leu Thr785 79la Leu Trp Tyr Ile Tyr Ser His Thr Gly Glu Thr Ala Gly Arg 88ln Val Pro Thr Ser Pro Pro Ala Ser Glu Asn Ser Ser Ala Ala823r Ile Gly Ser Thr Gln Ser Thr Pro Cys Ser Ser Ser Ser Thr 835 84a492omo sapiens 49ggaaagcatc caacggagtg gaagcagcat gaaggaagag ccgctgggca gcggcatgaa 6gcgg acgtggatgc agggcgccgg ggtgctggac gccaacacgg cggcgcagaggtgggt ctggcccggg ctcactttga gaagcagccg ccttccaatc tgcggaaatc ttcttc cacttcgtcc tggccctcta cgacagacag ggccagcccg tggagatcga 24agcg tttgtggggt tcgtggagaa ggaaaaagaa gccaacagcg aaaagaccaa 3gaatt cactaccgga ttcagcttct ctacagcaatgggataagga cggagcagga 36cgtg cgcctcattg actccatgac aaaacaagcc atagtgtatg aaggccaaga 42ccca gaaatgtgcc gagtcttgct cacacatgag atcatgtgca gccgctgttg 48gaaa agctgtggca accgaaatga gactccctca gatccagtga taattgacag 54cttg aaatttttcctcaaatgtaa ccaaaattgc ctaaagaatg cgggaaaccc 6acatg cggagattcc aggtcgtggt gtctacgaca gtcaatgtgg atggccatgt 66agct cctgataaca tgtttgtcca taataattcc aagcatgggc ggagggctcg 72tgac ccctcggaag gtacgccctc ttatctggaa catgctactc cctgtatcaa78cagc ccgagtgaag gatggacgac cggaggtgcg actgtcatca tcatagggga 84cttt gatgggttac aggtcatatt cggtaccatg ctggtctgga gtgagttgac 9ctcat tccatccgtg tgcagacccc tcctcggcac atccctggtg ttgtggaagt 96gtcc tacaaatcta agcagttctg caaaggaacaccaggcagat tcatttatac gcgcaac gaacccacca tcgattatgg tttccagagg ttacagaagg tcattcctcg ccctggt gaccctgagc gtttgccaaa ggaagtaata ctgaaaaggg ctgcggatct agaagca ctgtatggga tgccacacaa caaccaggaa atcattctga agagagcggc cattgccgaggccctgt acagtgtccc ccgcaaccac aaccaactcc cggcccttgc cacctcg gtccacgcag ggatgatggg cgtgaattcg ttcagtggac aactggccgt tgtctcc gaggcatcac aagccaccaa tcagggtttc acccgcaact caagcagcgt accacac gggtacgtgc cgagcaccac tccccagcag accaactataactccgtcac gagcatg aacggatacg gctctgccgc aatgtccaat ttgggtggct cccccacctt caacggc tcagctgcca actcccccta tgccatagtg ccatccagcc ccaccatggc ctccaca agcctcccct ccaactgcag cagctcctcg ggcatcttct ccttctcacc caacatg gtctcagccgtgaaacagaa gagtgctttc gcaccagtcg tcagacccca ctcccca cctcccacct gcaccagcac caacgggaac agcctgcaag cgatatctgg gattgtt cctcctatgt gaaagaattg ccttgaagaa ttgtattaat gaagaggttg tctgcta cagagagtaa tctgatacaa gtcccagagt ggaactttta actcaggccttaagagg aatcacacaa taactgcaga tttttaaaca aaatcaccga ccttgcaaat gaaattg gaagaggaat ctgaaagtgc agggtgttgg ttaaagttgt acctcccaag tttgggg atatatttat tctgtattga caaaagcaaa tccacttttt ctttttcttt 2ttttta agcttaattc tgcaatcatttgacttttat ataccgtaat gctctataca 2acacta taaataagac tccatgtttt aatttatgtt tttaaagctg tgtaaaggaa 2gaagtg gtgatattta c 26PRThomo sapiens 5r Ile Gln Arg Ser Gly Ser Ser Met Lys Glu Glu Pro Leu Glyly Met Asn Ala ValArg Thr Trp Met Gln Gly Ala Gly Val Leu 2Asp Ala Asn Thr Ala Ala Gln Ser Gly Val Gly Leu Ala Arg Ala His 35 4 Glu Lys Gln Pro Pro Ser Asn Leu Arg Lys Ser Asn Phe Phe His 5Phe Val Leu Ala Leu Tyr Asp Arg Gln Gly Gln Pro Val Glu IleGlu65 7Arg Thr Ala Phe Val Gly Phe Val Glu Lys Glu Lys Glu Ala Asn Ser 85 9 Lys Thr Asn Asn Gly Ile His Tyr Arg Ile Gln Leu Leu Tyr Ser Gly Ile Arg Thr Glu Gln Asp Phe Tyr Val Arg Leu Ile Asp Ser Thr Lys GlnAla Ile Val Tyr Glu Gly Gln Asp Lys Ser Pro Glu Cys Arg Val Leu Leu Thr His Glu Ile Met Cys Ser Arg Cys Cys Asp Lys Lys Ser Cys Gly Asn Arg Asn Glu Thr Pro Ser Asp Pro Val Ile Asp Arg Phe Phe Leu Lys Phe PheLeu Lys Cys Asn Gln Asn Leu Lys Asn Ala Gly Asn Pro Arg Asp Met Arg Arg Phe Gln Val 2al Ser Thr Thr Val Asn Val Asp Gly His Val Leu Ala Ala Pro 222n Met Phe Val His Asn Asn Ser Lys His Gly Arg Arg Ala Arg225234u Asp Pro Ser Glu Gly Thr Pro Ser Tyr Leu Glu His Ala Thr 245 25o Cys Ile Lys Ala Ile Ser Pro Ser Glu Gly Trp Thr Thr Gly Gly 267r Val Ile Ile Ile Gly Asp Asn Phe Phe Asp Gly Leu Gln Val 275 28e Phe Gly ThrMet Leu Val Trp Ser Glu Leu Thr Gly Pro His Ser 29rg Val Gln Thr Pro Pro Arg His Ile Pro Gly Val Val Glu Val33hr Leu Ser Tyr Lys Ser Lys Gln Phe Cys Lys Gly Thr Pro Gly Arg 325 33e Ile Tyr Thr Ala Arg Asn Glu Pro ThrIle Asp Tyr Gly Phe Gln 345u Gln Lys Val Ile Pro Arg His Pro Gly Asp Pro Glu Arg Leu 355 36o Lys Glu Val Ile Leu Lys Arg Ala Ala Asp Leu Val Glu Ala Leu 378y Met Pro His Asn Asn Gln Glu Ile Ile Leu Lys Arg Ala Ala38539le Ala Glu Ala Leu Tyr Ser Val Pro Arg Asn His Asn Gln Leu 44la Leu Ala Asn Thr Ser Val His Ala Gly Met Met Gly Val Asn 423e Ser Gly Gln Leu Ala Val Asn Val Ser Glu Ala Ser Gln Ala 435 44r Asn Gln GlyPhe Thr Arg Asn Ser Ser Ser Val Ser Pro His Gly 456l Pro Ser Thr Thr Pro Gln Gln Thr Asn Tyr Asn Ser Val Thr465 478r Met Asn Gly Tyr Gly Ser Ala Ala Met Ser Asn Leu Gly Gly 485 49r Pro Thr Phe Leu Asn Gly Ser Ala AlaAsn Ser Pro Tyr Ala Ile 55ro Ser Ser Pro Thr Met Ala Ser Ser Thr Ser Leu Pro Ser Asn 5525Cys Ser Ser Ser Ser Gly Ile Phe Ser Phe Ser Pro Ala Asn Met Val 534a Val Lys Gln Lys Ser Ala Phe Ala Pro Val Val Arg Pro Gln545556r Pro Pro Pro Thr Cys Thr Ser Thr Asn Gly Asn Ser Leu Gln 565 57a Ile Ser Gly Met Ile Val Pro Pro Met 58homo sapiens 5cccg cggtgcggag tatggggcgc tgatggccat ggagggctac tggcgcttcc 6tgct ggggtcggcactgctcgtcg gcttcctgtc ggtgatcttc gccctcgtct cctcca ctaccgagag gggcttggct gggatgggag cgcactagag tttaactggc agtgct catggtcacc ggcttcgtct tcatccaggg catcgccatc atcgtctaca 24cgtg gacctggaaa tgcagcaagc tcctgatgaa atccatccat gcagggttaa3gttgc tgccattctt gcaattatct ctgtggtggc cgtgtttgag aaccacaatg 36atat agccaatatg tacagtctgc acagctgggt tggactgata gctgtcatat 42tgtt acagcttctt tcaggttttt cagtctttct gcttccatgg gctccgcttt 48gagc atttctcatg cccatacatg tttattctggaattgtcatc tttggaacag 54caac agcacttatg ggattgacag agaaactgat tttttccctg agagatcctg 6agtac attcccgcca gaaggtgttt tcgtaaatac gcttggcctt ctgatcctgg 66gggc cctcattttt tggatagtca ccagaccgca atggaaacgt cctaaggagc 72ctac cattcttcatccaaatggag gcactgaaca gggagcaaga ggttccatgc 78actc tggcaacaac atggacaaat cagattcaga gttaaacagt gaagtagcag 84aaag aaacttagct ctggatgagg ctgggcagag atctaccatg taaaatgttg 9ataga gccatataac gtcacgtttc aaaactagct ctacagtttt gcttctccta96atat gataattggg ctatgtagta tcaatattta ctttaatcac aaaggatggt ttgaaat aatttgtatt gattgaggcc tatgaactga cctgaattgg aaaggatgtg aatataa ataatagcag atataaatta aaaaaaaaaa aaaagaaaa 86PRThomo sapiens 52Met Ala Met Glu Gly Tyr TrpArg Phe Leu Ala Leu Leu Gly Ser Alaeu Val Gly Phe Leu Ser Val Ile Phe Ala Leu Val Trp Val Leu 2His Tyr Arg Glu Gly Leu Gly Trp Asp Gly Ser Ala Leu Glu Phe Asn 35 4 His Pro Val Leu Met Val Thr Gly Phe Val Phe Ile Gln Gly Ile 5Ala Ile Ile Val Tyr Arg Leu Pro Trp Thr Trp Lys Cys Ser Lys Leu65 7Leu Met Lys Ser Ile His Ala Gly Leu Asn Ala Val Ala Ala Ile Leu 85 9 Ile Ile Ser Val Val Ala Val Phe Glu Asn His Asn Val Asn Asn Ala Asn Met Tyr Ser LeuHis Ser Trp Val Gly Leu Ile Ala Val Cys Tyr Leu Leu Gln Leu Leu Ser Gly Phe Ser Val Phe Leu Leu Trp Ala Pro Leu Ser Leu Arg Ala Phe Leu Met Pro Ile His Val Tyr Ser Gly Ile Val Ile Phe Gly Thr Val Ile Ala ThrAla Leu Met Leu Thr Glu Lys Leu Ile Phe Ser Leu Arg Asp Pro Ala Tyr Ser Phe Pro Pro Glu Gly Val Phe Val Asn Thr Leu Gly Leu Leu Ile 2al Phe Gly Ala Leu Ile Phe Trp Ile Val Thr Arg Pro Gln Trp 222g Pro Lys Glu Pro Asn Ser Thr Ile Leu His Pro Asn Gly Gly225 234u Gln Gly Ala Arg Gly Ser Met Pro Ala Tyr Ser Gly Asn Asn 245 25t Asp Lys Ser Asp Ser Glu Leu Asn Ser Glu Val Ala Ala Arg Lys 267n Leu Ala Leu Asp GluAla Gly Gln Arg Ser Thr Met 275 282655DNAhomo sapiens 53cgcctagccg cgccggtccc agaagtggcg aaagccgcag ccgagtccag gtcacgccga 6tgcc cttttaaggg ggagccttga aacggcgcct gggttccatg tttgcatccg gcggga aggaaactcc atgttgtaac aaagtttcct ccgcgccccctccctccccc ccctag aacctggctc ccctcccctc cggagctcgc ggggatccct ccctcccacc 24ctcc cccccgcgcc ccgattccgg ccccagccgg gggggaggcc gggcgcccgg 3agtcc ggccggagcg gagcgcgccc ggccccatgg acagctcggc cgtcattact 36agca aggaggaggc tcggggcccgctgcggggca aaggtgacca gaagtcagca 42caga agccccgaag ccggggcatc ctccactcac tcttctgctg tgtctgccgg 48gggg aggccctgcc tgctcacagc ggggcgcccc tgcttgtgga ggagaatgga 54ccta agcagacccc agtccaatac ctgctccctg aggccaaggc ccaggactca 6gatctgcgtggtcat cgacctggac gagaccctgg tgcacagctc cttcaagcca 66aacg cggacttcat catccctgtg gagattgatg gggtggtcca ccaggtctac 72aagc gtcctcacgt ggatgagttc ctgcagcgaa tgggcgagct ctttgaatgt 78ttca ctgctagcct cgccaagtac gcagacccag tagctgacctgctggacaaa 84gcct tccgggcccg gctgtttcga gagtcctgcg tcttccaccg ggggaactac 9ggacc tgagccggtt gggtcgagac ctgcggcggg tgctcatcct ggacaattca 96tcct atgtcttcca tccagacaat gctgtaccgg tggcctcgtg gtttgacaac agtgaca cagagctcca cgacctcctccccttcttcg agcaactcag ccgtgtggac gtgtact cagtgctcag gcagccacgg ccagggagct agtgagggtg atggggccag ctgcccc tgaccaatga tacccacacc tcctcccagg aagactgccc aggcctttgt gaaaacc catgggccgc cgccacactc agtgccatgg ggaagcgggc gtctcccccagccccac caggcggtgt aggggcagca ggctgcactg aggaccgtga gctccaggcc tgtcagt gccttcaaac ctcctcccct attctcaggg gacctggggg gccctgcctg ctccctt tttctgtctc tgtccatgct gccatgtttc tctgctgcca aattgggccc ggcccct tccggttctg cttcctgggggcagggttcc tgccttggac ccccagtctg acggtgg acatcaagtg ccttgcatag agccccctct tccccgccca gctttcccag cacagct ctaggctggg aggggagaac cagcccctcc ccctgcccca cctcctccct gactgag agggccccta ccaacctttg cctctgcctt ggagggaggg gaggtctgttactgggg aaggcagcag gagtctgtcc ttcaggcccc acagtgcagc ttctccaggg acagctg agggctgctc cctgcatcat ccaagcaatg acctcagact tctgccttaa gccccgg ggcttggctc ccccagctct gagcgtgggg gcataggcag gacccccctt gtgccat ataaatatgt acatgtgtatatagattttt aggggaagga gagagggaag cagggta gagacacccc tcccttgccc ctttcctggg cccagaagtt ggggggaggg gaaagga tttttacatt ttttaaactg ctattttctg aatggaacaa gctgggccaa 2cccagg ccctgtcctc tgtccctcac acccctttgc tccgttcatt cattcaaaaa2tttctt gagcaccttc tgtgcccagc atatgctagg cccaccagct aagtgtgtgt 2ggtctc tacgccagct catcagtgcc tccttgccca tccttcaccg gtgcctttgg 222tgta ggaggtggga ccttctgtgg ggtttgggga tctccaggaa gcccgaccaa 228ccct tcccctgtgc caacccatctcctacagccc cctgcctgat cccctgctgg 234gcag ctcccaggat atcctgcctt ccaactgttt ctgaagcccc tcctcctaac 24gattc cggaggtcaa ggccttgggc tctccccagg gtctaacggt taaggggacc 246ccag tgccaagggg gatgtcaagt ggtgatgtcg ttgtgctccc ctcccccaga252gggc ggggggtgaa tatggttggc ctgcatcagg tggccttccc atttaagtgc 258tgtg actgagagcc ctagtgtgat gagaactaaa gagaaagcca gacccctaaa 264aaaa aaaaa 26555426o sapiens 54Met Asp Ser Ser Ala Val Ile Thr Gln Ile Ser Lys Glu Glu Ala Argro Leu Arg Gly Lys Gly Asp Gln Lys Ser Ala Ala Ser Gln Lys 2Pro Arg Ser Arg Gly Ile Leu His Ser Leu Phe Cys Cys Val Cys Arg 35 4 Asp Gly Glu Ala Leu Pro Ala His Ser Gly Ala Pro Leu Leu Val 5Glu Glu Asn Gly Ala Ile Pro LysGln Thr Pro Val Gln Tyr Leu Leu65 7Pro Glu Ala Lys Ala Gln Asp Ser Asp Lys Ile Cys Val Val Ile Asp 85 9 Asp Glu Thr Leu Val His Ser Ser Phe Lys Pro Val Asn Asn Ala Phe Ile Ile Pro Val Glu Ile Asp Gly Val Val His Gln Val Tyr Leu Lys Arg Pro His Val Asp Glu Phe Leu Gln Arg Met Gly Glu Phe Glu Cys Val Leu Phe Thr Ala Ser Leu Ala Lys Tyr Ala Asp Pro Val Ala Asp Leu Leu Asp Lys Trp Gly Ala Phe Arg Ala Arg Leu Arg Glu SerCys Val Phe His Arg Gly Asn Tyr Val Lys Asp Leu Arg Leu Gly Arg Asp Leu Arg Arg Val Leu Ile Leu Asp Asn Ser 2la Ser Tyr Val Phe His Pro Asp Asn Ala Val Pro Val Ala Ser 222e Asp Asn Met Ser Asp Thr Glu Leu HisAsp Leu Leu Pro Phe225 234u Gln Leu Ser Arg Val Asp Asp Val Tyr Ser Val Leu Arg Gln 245 25o Arg Pro Gly Ser 26DNAhomo sapiens 55gcccgagcga gggcgcttcg ctcccagcca ggacatggcc gcacctctcc gcatcaggag 6ctca cggacttctcgcccaactcc ctgagcgctc cctcgtttcg atctttagaa ccgctt tctttctggg gccgtgacga ggggcaggga gcggcgagca aggatgcgtt accgcg agggcgcgcg tctcgggtgc cgccgtgggt cccgacgcgg aagccgagcc 24gcct gcctcgactt ccccacagcg cttccgccgc cgcctgccgt gcttgtatgt3aagaa gccggacacc atgatcctaa cacaaattga agccaaggaa gcttgtgatt 36gggc aactggtttc ccccagtatg cacagcttta tgaagatttc ctgttcccca 42tttc cttggtcaag agagagcatg attttttgga cagagatgcc attgaggctc 48cgtc taaatacttt aaacaaatgt gcggtgatgaagctagaaat tagtcctcat 54cgag tgacgattca gacgaggatg agccttgtgc catcagtggc aaatggactt 6aggga cagcaagagg tggtcccggc ttgaagagtt tgatgtcttt tctccaaaac 66tggt ccctgggtcc ccagacgact cccacccgaa ggacggcccc agccccggag 72tgat ggacctcagcgagcgccagg aggtgtcttc cgtccgcagc ctcagcagca 78gcct ccccagccac gcgcccccca gcgaggatgc tgccaccccc cggactaact 84tcag cgtttgctcc tccagcaact tggcaggcaa

tgacgactct ttcggcagcc 9tctcc caaggaactg tccagcttca gcttcagcat gaaaggccac gaaaaaactg 96ccaa gacgcgcagt ctgctgaaac ggatggagag cctgaagctc aagagctccc acagcaa gcacaaagcg ccctcaaagc tggggttgat catcagcggg cccatcttgc aggggatggatgaggag aagctgaagc agctcaactg cgtggagatc tccgccctca gcaaccg catcaacgtc cccatggtac gaaagaggag cgtttccaac tccacgcaga gcagcag cagcagccag tcggagacca gcagcgcggt cagcacgccc agccctgtta ggacccg gagcctcagt gcgtgcaaca agcgggtggg catgtacttagagggcttcg ctttcaa tcagtcaaca tttaacaacg tgatggagca gaactttaag aaccgcgaga acccaga ggacacggtg ttctacatcc ctgaagatca caagcctggc actttcccca ctctcac caatggcagt ttctccccct cggggaataa cggctctgtg aactggagga gaagctt ccacggccctggccacatca gcctcaggag ggaaaacagt agcgacagcc aggaact gaagagacgc aattcttcca gctccatgag cagccgcctg agcatctacg acgtgcc gggctccatc ctctactcca gttcagggga cctggcggat ctggagaacg acatctt ccccgagctg gacgacatcc tctaccacgt gaaggggatg cagcggatagatcagtg gtcggagaag ttttctgatg agggagattc ggactcagcc ctggactcgg ctccctg cccgtcctct ccaaaacaga tacacctgga tgtggacaac gaccgaacca ccagcga cctggacagc acaggcaact ccctgaatga accggaagag ccctccgaga cggaaag aagggattct ggggttggggcttccctaac caggtccaac aggcaccgac gatggca cagtttccag agctcacatc ggccaagcct caactctgta tcactacaga 2ctgcca gtctgtggcc cagatgaacc tgctgcagaa atactcactc ctaaagctaa 2cctgct ggagaaatac acaccttcta acaagcatgg ttttagctgg gccgtgccca2catgaa gaggatcaag gttccagact acaaggaccg gagtgtgttt ggggtcccac 222tcaa cgtgcagcgc acaggacaac cgttgcctca gagcatccag caggccatgc 228tccg gaaccattgt ttggatcagg ttgggctctt cagaaaatcg ggggtcaagt 234ttca ggctctgcgc cagatgaatgaaggtgccat agactgtgtc aactacgaag 24tctgc ttatgacgtg gcagacatgc tgaagcagta ttttcgagat cttcctgagc 246tgac gaacaaactc tcagaaacct ttctacagat ctaccaatat gtgcccaagg 252gact gcaggccatc aaggctgcca tcatgctgct gcctgacgag aaccgggagg258agac cctgctttat ttcctgagcg atgtcacagc agccgtaaaa gaaaaccaga 264caac caacctggcc gtgtgcttag cgccttccct cttccatctc aacaccctga 27gagaa ttcctctccc aggggtaatg caaagaaaac aaagtttggg caaaccagat 276gatt tgaatgaaaa cctagctgccactcaagggc tggcccatat gatcgccgag 282aagc ttttccaggt tcccgaggaa atgagccgat gtcgtaattc ctataccgaa 288ctga agcccctcac tctggaagca ctcgggcacc tgggtaatga tgactcagct 294caac acttcctcca ggactgtgtg gatggcctgt ttaaagaagt caaagagaag3aaggct gggtcagcta ctccacttcg gagcaggctg agctgtccta taagaaggtg 3aaggac cccctctgag gctttggagg tcagtcattg aagtccctgc tgtgccagag 3tcttaa agcgcctact taaagaacag cacctctggg atgtagacct gttggattca 3tgatcg aaattctgga cagccaaactgaaatttacc agtatgtcca aaacagtatg 324catc ctgctcgaga ctacgttgtt ttaagaacct ggaggactaa tttacccaaa 33ctgtg cccttttact aacctctgtg gatcacgatc gcgcacctgt ggtgggtgtg 336aatg tgctcttgtc caggtatttg attgaaccct gtgggccagg aaaatccaaa342taca tgtgcagagt tgacttaagg ggccacatgc cagaatggta cacaaaatct 348catt tgtgtgcagc tgaagttgta aagatccggg attccttcag taaccagaac 354acca aagacaccaa atctaggtga tcactgaagc aacgcaaccg cttccaccac 36tgttt gtttctagaa cttttgccagtccttgaaga atgggttctg tgtctaatcc 366aaag aaaactacaa gctggagtgt aggaattgac tatagcaatt tgatacattt 372ctgc ttcctgtttg ttgagggtct gtattcatag accttgactg gaatatgtaa 378gcaa aaaaaaaaaa aaaa 38o sapiens 56Met Cys Arg Lys LysPro Asp Thr Met Ile Leu Thr Gln Ile Glu Alalu Ala Cys Asp Trp Leu Arg Ala Thr Gly Phe Pro Gln Tyr Ala 2Gln Leu Tyr Glu Asp Phe Leu Phe Pro Ile Asp Ile Ser Leu Val Lys 35 4 Glu His Asp Phe Leu Asp Arg Asp Ala Ile Glu Ala LeuCys Arg 5Arg Leu Asn Thr Leu Asn Lys Cys Ala Val Met Lys Leu Glu Ile Ser65 7Pro His Arg Lys Arg Ser Asp Asp Ser Asp Glu Asp Glu Pro Cys Ala 85 9 Ser Gly Lys Trp Thr Phe Gln Arg Asp Ser Lys Arg Trp Ser Arg Glu Glu PheAsp Val Phe Ser Pro Lys Gln Asp Leu Val Pro Gly Pro Asp Asp Ser His Pro Lys Asp Gly Pro Ser Pro Gly Gly Thr Met Asp Leu Ser Glu Arg Gln Glu Val Ser Ser Val Arg Ser Leu Ser Ser Thr Gly Ser Leu Pro Ser His AlaPro Pro Ser Glu Asp Ala Thr Pro Arg Thr Asn Ser Val Ile Ser Val Cys Ser Ser Ser Asn Ala Gly Asn Asp Asp Ser Phe Gly Ser Leu Pro Ser Pro Lys Glu 2er Ser Phe Ser Phe Ser Met Lys Gly His Glu Lys Thr Ala Lys 222s Thr Arg Ser Leu Leu Lys Arg Met Glu Ser Leu Lys Leu Lys225 234r His His Ser Lys His Lys Ala Pro Ser Lys Leu Gly Leu Ile 245 25e Ser Gly Pro Ile Leu Gln Glu Gly Met Asp Glu Glu Lys Leu Lys 267u Ser Cys ValGlu Ile Ser Ala Leu Asn Gly Asn Arg Ile Asn 275 28l Pro Met Val Arg Lys Arg Ser Val Ser Asn Ser Thr Gln Thr Ser 29er Ser Ser Gln Ser Glu Thr Ser Ser Ala Val Ser Thr Pro Ser33ro Val Thr Arg Thr Arg Ser Leu Ser Ala CysAsn Lys Arg Val Gly 325 33t Tyr Leu Glu Gly Phe Asp Pro Phe Asn Gln Ser Thr Phe Asn Asn 345l Glu Gln Asn Phe Lys Asn Arg Glu Ser Tyr Pro Glu Asp Thr 355 36l Phe Tyr Ile Pro Glu Asp His Lys Pro Gly Thr Phe Pro Lys Ala 378r Asn Gly Ser Phe Ser Pro Ser Gly Asn Asn Gly Ser Val Asn385 39rg Thr Gly Ser Phe His Gly Pro Gly His Ile Ser Leu Arg Arg 44sn Ser Ser Asp Ser Pro Lys Glu Leu Lys Arg Arg Asn Ser Ser 423r Met Ser Ser ArgLeu Ser Ile Tyr Asp Asn Val Pro Gly Ser 435 44e Leu Tyr Ser Ser Ser Gly Asp Leu Ala Asp Leu Glu Asn Glu Asp 456e Pro Glu Leu Asp Asp Ile Leu Tyr His Val Lys Gly Met Gln465 478e Val Asn Gln Trp Ser Glu Lys Phe Ser AspGlu Gly Asp Ser 485 49p Ser Ala Leu Asp Ser Val Ser Pro Cys Pro Ser Ser Pro Lys Gln 55is Leu Asp Val Asp Asn Asp Arg Thr Thr Pro Ser Asp Leu Asp 5525Ser Thr Gly Asn Ser Leu Asn Glu Pro Glu Glu Pro Ser Glu Ile Pro 534g Arg Asp Ser Gly Val Gly Ala Ser Leu Thr Arg Ser Asn Arg545 556g Leu Arg Trp His Ser Phe Gln Ser Ser His Arg Pro Ser Leu 565 57n Ser Val Ser Leu Gln Ile Asn Cys Gln Ser Val Ala Gln Met Asn 589u Gln Lys Tyr SerLeu Leu Lys Leu Thr Ala Leu Leu Glu Lys 595 6yr Thr Pro Ser Asn Lys His Gly Phe Ser Trp Ala Val Pro Lys Phe 662s Arg Ile Lys Val Pro Asp Tyr Lys Asp Arg Ser Val Phe Gly625 634o Leu Thr Val Asn Val Gln Arg Thr Gly GlnPro Leu Pro Gln 645 65r Ile Gln Gln Ala Met Arg Tyr Leu Arg Asn His Cys Leu Asp Gln 667y Leu Phe Lys Lys Ser Gly Val Lys Ser Arg Ile Gln Ala Leu 675 68g Gln Met Asn Glu Gly Ala Ile Asp Cys Val Asn Tyr Glu Gly Gln 69la Tyr Asp Val Ala Asp Met Leu Lys Gln Tyr Phe Arg Asp Leu77ro Glu Pro Leu Met Thr Asn Lys Leu Ser Glu Thr Phe Leu Gln Ile 725 73r Gln Tyr Val Pro Lys Asp Gln Arg Leu Gln Ala Ile Lys Ala Ala 745t Leu Leu Pro AspGlu Asn Arg Val Val Leu Gln Thr Leu Leu 755 76r Phe Leu Cys Asp Val Thr Ala Ala Val Lys Glu Asn Gln Met Thr 778r Asn Leu Ala Val Cys Leu Ala Pro Ser Leu Phe His Leu Asn785 79eu Lys Arg Glu Asn Ser Ser Pro Arg Val MetGln Arg Lys Gln 88eu Gly Lys Pro Asp Gln Lys Asp Leu Asn Glu Asn Leu Ala Ala 823n Gly Leu Ala His Met Ile Ala Glu Cys Lys Lys Leu Phe Gln 835 84l Pro Glu Glu Met Ser Arg Cys Arg Asn Ser Tyr Thr Glu Gln Glu 856s Pro Leu Thr Leu Glu Ala Leu Gly His Leu Gly Asn Asp Asp865 878a Asp Tyr Gln His Phe Leu Gln Asp Cys Val Asp Gly Leu Phe 885 89s Glu Val Lys Glu Lys Phe Lys Gly Trp Val Ser Tyr Ser Thr Ser 99ln Ala Glu Leu SerTyr Lys Lys Val Ser Glu Gly Pro Arg Leu 9925Arg Leu Trp Arg Ser Val Ile Glu Val Pro Ala Val Pro Glu Glu Ile 934s Arg Leu Leu Lys Glu Gln His Leu Trp Asp Val Asp Leu Leu945 956r Lys Val Ile Glu Ile Leu Asp Ser Gln ThrGlu Ile Tyr Gln 965 97r Val Gln Asn Ser Met Ala Pro His Pro Ala Arg Asp Tyr Val Val 989g Thr Trp Arg Thr Asn Leu Pro Lys Gly Ala Cys Ala Leu Leu 995 hr Ser Val Asp His Asp Arg Ala Pro Val Val Gly Val Arg Val Asn Val Leu Leu Ser Arg Tyr Leu Ile Glu Pro Cys Gly Pro 3ly Lys Ser Lys Leu Thr Tyr Met Cys Arg Val Asp Leu Arg Gly 45 Met Pro Glu Trp Tyr Thr Lys Ser Phe Gly His Leu Cys Ala 6la Glu Val Val Lys Ile ArgAsp Ser Phe Ser Asn Gln Asn Thr 75 Thr Lys Asp Thr Lys Ser Arg 9DNAhomo sapiens 57ctggccccga gcagctgaag cctggggtca gcaggcgctg cgggcgcagc tccggtgcaa 6acac gacacatgca gtggcttctg gactgcgcga tgactggacg caagtaacttgtctgc agacaagagg aagagaagat gaaggaagac tgtctgccga gttctcacgt atcagt gacagcaagt ccattcagaa gtcggagctc ttaggcctgc tgaaaaccta 24ctac catgagggca agagcttcca gctgagacac cgtgaggaag aagggactct 3tcgag gggctcctca acattgcctg ggggctgaggcggcccatcc ggctgcagat 36tgac cgggagcagg tgcacctccc ctccacctca tggatgccca gacggcctag 42tcta aaggagccat cgccccagaa cgggaacatc acagcccagg ggccaagcat 48agtg cacaaggctg agagttccac agacagctcg gggcccctgg aggaggcaga 54cccc cagctgatgcggaccaagag cgacgccagt tgcatgagcc agaggaggcc 6gccgc gcccccggtg aggcccagcg catccggcga caccggttct ctatcaacgg 66ctac aatcataaga cctccgtgtt tactccagcc tatggatccg tgaccaatgt 72caac agcaccatga caaccctgca ggtgctcacc ctgctgctga acaaatttag78agat ggccccagtg agttcgcact ctacatcgtt cacgagtctg gggagcggac 84aaaa gactgcgagt acccgctgat ttccagaatc ctgcatgggc catgtgagaa 9ccagg atcttcctga tggaagctga cttgggcgtg gaagtccccc atgaagtcgc 96catt aagtttgaaa tgccggtgct ggacagttttgttgaaaaat taaaagaaga agaaaga gaaataatca aactgaccat gaagttccaa gccctgcgtc tgacgatgct gcgcctg gagcagctgg tggaggccaa gtaactggcc aacacctgcc tcttccaaag ccagcag tggcaggtgt acactgagcc ctggttgctg gccccggccg gtcacattga atggccaccgcctgacg aatcgagtgc ctgtgtgtct acctctctga agcctgagca tgattcc cacagccagc tcttggctcc aagatgagca cccacaggaa gccgacccag tgagggg ccaggaactt gctgggtcag atctgtgtgg ccagccctgt ccacaccatg ctcctgc actggagagc agtgctggcc cagcccctgc ggcttaggcttcatctgctt cattgcc tgtcccagag cccctgtggg tccacaagcc cctgtcctct tccttcatat attcttg tctgccctca tatcacgctg ccccacagga atgctgctgg gaaaagcagg tgccagc aggtatgaga tctagcctgc tttcagccat caccttgcca cagtgtcccc ttctaag cctccaatatcaccctgtga gcctcgcaca gctcagcccc aacacagagg gaccagg aataaggcca caagtatctc actttctcgc agggaatcaa tcttagcttc agagaga cttaaagcga ttctgacaag gagctgctgc aagaaacgcg gtcattcaat attgagg agggtccaca tggcattgag agggtgctgc ccgctcaatg cccagcagcactggaag gcagtgctca gccccatcac cactgtcccg tggatgcctg tgtacctctt ttttctg ggcttgcgtt tctctcctct agtgggtggg gatgactttc aatgactttc acttccc ctgaaggaag aatgataagg agaaatgtct gttttgagga aagggctttg 2ccccag atactgaaca atttgtgtttgtgactgatg gagaatttca ggaatgaatg 2agcctt tgcgaaacta tgcaacagtt tacatcagtc atgtgaagta tttgtctaaa 2agcaaa ctgaagacca aattattctc ctgttgaggt ccgtggatgg cagatttaaa 222aacc acaaaggctt gcaaagatag gagaggctcc atctctaatg catgtagaag228acgg gtgcccatca agagcatagc ttggaagcca ccatgctgtg cggaactgcg 234caaa tgtcacagca ggatttcccc aacccagctc catcatcaca gacacagaga 24agggg aggcctgccc actgttttgt cgactctgcc ctcctctggc agcatagatc 246tgct caataaaggt gtgctgtattgaactgaaga a 25PRThomo sapiens 58Met Lys Glu Asp Cys Leu Pro Ser Ser His Val Pro Ile Ser Asp Serer Ile Gln Lys Ser Glu Leu Leu Gly Leu Leu Lys Thr Tyr Asn 2Cys Tyr His Glu Gly Lys Ser Phe Gln Leu Arg His Arg Glu Glu Glu 35 4 Thr Leu Ile Ile Glu Gly Leu Leu Asn Ile Ala Trp Gly Leu Arg 5Arg Pro Ile Arg Leu Gln Met Gln Asp Asp Arg Glu Gln Val His Leu65 7Pro Ser Thr Ser Trp Met Pro Arg Arg Pro Ser Cys Pro Leu Lys Glu 85 9 Ser Pro Gln Asn Gly Asn IleThr Ala Lys Gly Pro Ser Ile Gln Val His Lys Ala Glu Ser Ser Thr Asp Ser Ser Gly Pro Leu Glu Ala Glu Glu Ala Pro Gln Leu Met Arg Thr Lys Ser Asp Ala Ser Met Ser Gln Arg Arg Pro Lys Cys Arg Ala Pro Gly Glu AlaGln Arg Ile Arg Arg His Arg Phe Ser Ile Asn Gly His Phe Tyr Asn His Thr Ser Val Phe Thr Pro Ala Tyr Gly Ser Val Thr Asn Val Arg Asn Ser Thr Met Thr Thr Leu Gln Val Leu Thr Leu Leu Leu Asn 2he ArgVal Glu Asp Gly Pro Ser Glu Phe Ala Leu Tyr Ile Val 222u Ser Gly Glu Arg Thr Lys Leu Lys Asp Cys Glu Tyr Pro Leu225 234r Arg Ile Leu His Gly Pro Cys Glu Lys Ile Ala Arg Ile Phe 245 25u Met Glu Ala Asp Leu Gly Val GluVal Pro His Glu Val Ala Gln 267e Lys Phe Glu Met Pro Val Leu Asp Ser Phe Val Glu Lys Leu 275 28s Glu Glu Glu Glu Arg Glu Ile Ile Lys Leu Thr Met Lys Phe Gln 29eu Arg Leu Thr Met Leu Gln Arg Leu Glu Gln Leu Val GluAla33ys59homo sapiens 59atgagtgcac ttttccttgg tgtgggagtg agggcagagg aagctggagc gagggtgcaa 6gttc caagtgggac agatactgga gatcctcaaa gtaagcccct cggtgactgg ctggca ccatggaccc agagagcagt atctttattg aggatgccat taagtatttcaaaaag tgagcacaca gaatctgcta ctcctgctga ctgataatga ggcctggaac 24gtgg ctgctgctga actgcccagg aatgaggcag atgagctccg taaagctctg 3ccttg caagacaaat gatcatgaaa gacaaaaact ggcacgataa aggccagcag 36aact ggtttctgaa agagtttcct cggttgaaaagtgagcttga ggataacata 42ctcc gtgcccttgc agatggggtt cagaaggtcc acaaaggcac caccatcgcc 48gtgt ctggctctct cagcatttcc tctggcatcc tgaccctcgt cggcatgggt 54ccct tcacagaggg aggcagcctt gtactcttgg aacctgggat ggagttggga 6agccg ctttgaccgggattaccagc agtaccatgg actacggaaa gaagtggtgg 66gccc aagcccacga cctggtcatc aaaagccttg acaaattgaa ggaggtgagg 72ttgg gtgagaacat atccaacttt ctttccttag ctggcaatac ttaccaactc 78ggca ttgggaagga catccgtgcc ctcagacgag ccagagccaa tcttcagtca84catg cctcagcctc acgcccccgg gtcactgagc caatctcagc

tgaaagcggt 9ggtgg agagggttaa tgaacccagc atcctggaaa tgagcagagg agtcaagctc 96gtgg cccctgtaag cttctttctt gtgctggatg tagtctacct cgtgtacgaa aagcact tacatgaggg ggcaaagtca gagacagctg aggagctgaa gaaggtggct gagctgg aggagaagctaaacattctc aacaataatt ataagattct gcaggcggac gaactgt ga 83PRThomo sapiens 6r Ala Leu Phe Leu Gly Val Gly Val Arg Ala Glu Glu Ala Glyrg Val Gln Gln Asn Val Pro Ser Gly Thr Asp Thr Gly Asp Pro 2Gln Ser Lys Pro LeuGly Asp Trp Ala Ala Gly Thr Met Asp Pro Glu 35 4 Ser Ile Phe Ile Glu Asp Ala Ile Lys Tyr Phe Lys Glu Lys Val 5Ser Thr Gln Asn Leu Leu Leu Leu Leu Thr Asp Asn Glu Ala Trp Asn65 7Gly Phe Val Ala Ala Ala Glu Leu Pro Arg Asn Glu Ala AspGlu Leu 85 9 Lys Ala Leu Asp Asn Leu Ala Arg Gln Met Ile Met Lys Asp Lys Trp His Asp Lys Gly Gln Gln Tyr Arg Asn Trp Phe Leu Lys Glu Pro Arg Leu Lys Ser Glu Leu Glu Asp Asn Ile Arg Arg Leu Arg Leu AlaAsp Gly Val Gln Lys Val His Lys Gly Thr Thr Ile Ala Asn Val Val Ser Gly Ser Leu Ser Ile Ser Ser Gly Ile Leu Thr Leu Gly Met Gly Leu Ala Pro Phe Thr Glu Gly Gly Ser Leu Val Leu Glu Pro Gly Met Glu Leu Gly IleThr Ala Ala Leu Thr Gly Ile 2er Ser Thr Met Asp Tyr Gly Lys Lys Trp Trp Thr Gln Ala Gln 222s Asp Leu Val Ile Lys Ser Leu Asp Lys Leu Lys Glu Val Arg225 234e Leu Gly Glu Asn Ile Ser Asn Phe Leu Ser Leu Ala GlyAsn 245 25r Tyr Gln Leu Thr Arg Gly Ile Gly Lys Asp Ile Arg Ala Leu Arg 267a Arg Ala Asn Leu Gln Ser Val Pro His Ala Ser Ala Ser Arg 275 28o Arg Val Thr Glu Pro Ile Ser Ala Glu Ser Gly Glu Gln Val Glu 29al AsnGlu Pro Ser Ile Leu Glu Met Ser Arg Gly Val Lys Leu33hr Asp Val Ala Pro Val Ser Phe Phe Leu Val Leu Asp Val Val Tyr 325 33u Val Tyr Glu Ser Lys His Leu His Glu Gly Ala Lys Ser Glu Thr 345u Glu Leu Lys Lys Val Ala GlnGlu Leu Glu Glu Lys Leu Asn 355 36e Leu Asn Asn Asn Tyr Lys Ile Leu Gln Ala Asp Gln Glu Leu 378DNAhomo sapiens 6ttag agatccacat gtcagacccc atgtgcctca tcgagaactt taatgagcag 6gtta atcaggaagc tttggagatc ctgtctgccattacgcaacc tgtagttgtg cgattg tgggcctcta tcgcactggc aaatcctacc tgatgaacaa gctggctggg acaagg gcttctctgt tgcatctacg gtgcagtctc acaccaaggg aatttggata 24gtgc ctcatcccaa ctggccaaat cacacattag ttctgcttga caccgagggc 3agatg tagagaaggctgacaacaag aatgatatcc agatctttgc actggcactc 36agca gcacctttgt gtacaatact gtgaacaaaa ttgatcaggg tgctatcgac 42caca atgtgacaga actgacagat ctgctcaagg caagaaactc acccgacctt 48gttg aagatcctgc tgactctgcg agcttcttcc cagacttagt gtggactctg54ttct gcttaggcct ggaaatagat gggcaacttg tcacaccaga tgaatacctg 6ttccc taaggccaaa gcaaggtagt gatcaaagag ttcaaaattt caatttgcct 66tgta tacagaagtt ctttccaaaa aagaaatgct ttatctttga cttacctgct 72aaaa agcttgccca acttgaaaca ctgcctgatgatgagctaga gcctgaattt 78caag tgacagaatt ctgttcctac atctttagcc attctatgac caagactctt 84ggca tcatggtcaa tggatctcgt ctaaagaacc tggtgctgac ctatgtcaat 9cagca gtggggatct gccttgcata gagaatgcag tcctggcctt ggctcagaga 96tcag ctgcagtgcaaaaggccatt gcccactatg accagcaaat gggccagaaa cagctgc ccatggaaac cctccaggag ctgctggacc tgcacaggac cagtgagagg gccattg aagtcttcat gaaaaactct ttcaaggatg tagaccaaag tttccagaaa ttggaga ctctactaga tgcaaaacag aatgacattt gtaaacggaa cctggaagcatcggatt attgctcggc tttacttaag gatatttttg gtcctctaga agaagcagtg cagggaa tttattctaa gccaggaggc cataatctct tcattcagaa aacagaagaa aaggcaa agtactatcg ggagcctcgg aaaggaatac aggctgaaga agttctgcag tatttaa agtccaagga gtctgtgagtcatgcaatat tacagactga ccaggctctc gagacgg aaaaaaagaa gaaagaggca caagtgaaag cagaagctga aaaggctgaa caaaggt tggcggcgat tcaaaggcag aacgagcaaa tgatgcagga gagggagaga catcagg aacaagtgag acaaatggag atagccaaac aaaattggct ggcagagcaaaaaatgc aggaacaaca gatgcaggaa caggctgcac agctcagcac aacattccaa caaaata gaagccttct cagtgagctc cagcacgccc agaggactgt taataacgat ccatgtg ttttactcta a 86PRThomo sapiens 62Met Ala Leu Glu Ile His Met Ser Asp Pro Met Cys Leu Ile GluAsnsn Glu Gln Leu Lys Val Asn Gln Glu Ala Leu Glu Ile Leu Ser 2Ala Ile Thr Gln Pro Val Val Val Val Ala Ile Val Gly Leu Tyr Arg 35 4 Gly Lys Ser Tyr Leu Met Asn Lys Leu Ala Gly Lys Asn Lys Gly 5Phe Ser Val Ala Ser ThrVal Gln Ser His Thr Lys Gly Ile Trp Ile65 7Trp Cys Val Pro His Pro Asn Trp Pro Asn His Thr Leu Val Leu Leu 85 9 Thr Glu Gly Leu Gly Asp Val Glu Lys Ala Asp Asn Lys Asn Asp Gln Ile Phe Ala Leu Ala Leu Leu Leu Ser Ser Thr PheVal Tyr Thr Val Asn Lys Ile Asp Gln Gly Ala Ile Asp Leu Leu His Asn Thr Glu Leu Thr Asp Leu Leu Lys Ala Arg Asn Ser Pro Asp Leu Asp Arg Val Glu Asp Pro Ala Asp Ser Ala Ser Phe Phe Pro Asp Leu TrpThr Leu Arg Asp Phe Cys Leu Gly Leu Glu Ile Asp Gly Gln Val Thr Pro Asp Glu Tyr Leu Glu Asn Ser Leu Arg Pro Lys Gln 2er Asp Gln Arg Val Gln Asn Phe Asn Leu Pro Arg Leu Cys Ile 222s Phe Phe Pro Lys Lys Lys CysPhe Ile Phe Asp Leu Pro Ala225 234n Lys Lys Leu Ala Gln Leu Glu Thr Leu Pro Asp Asp Glu Leu 245 25u Pro Glu Phe Val Gln Gln Val Thr Glu Phe Cys Ser Tyr Ile Phe 267s Ser Met Thr Lys Thr Leu Pro Gly Gly Ile Met Val AsnGly 275 28r Arg Leu Lys Asn Leu Val Leu Thr Tyr Val Asn Ala Ile Ser Ser 29sp Leu Pro Cys Ile Glu Asn Ala Val Leu Ala Leu Ala Gln Arg33lu Asn Ser Ala Ala Val Gln Lys Ala Ile Ala His Tyr Asp Gln Gln 325 33t Gly GlnLys Val Gln Leu Pro Met Glu Thr Leu Gln Glu Leu Leu 345u His Arg Thr Ser Glu Arg Glu Ala Ile Glu Val Phe Met Lys 355 36n Ser Phe Lys Asp Val Asp Gln Ser Phe Gln Lys Glu Leu Glu Thr 378u Asp Ala Lys Gln Asn Asp Ile CysLys Arg Asn Leu Glu Ala385 39er Asp Tyr Cys Ser Ala Leu Leu Lys Asp Ile Phe Gly Pro Leu 44lu Ala Val Lys Gln Gly Ile Tyr Ser Lys Pro Gly Gly His Asn 423e Ile Gln Lys Thr Glu Glu Leu Lys Ala Lys Tyr Tyr Arg Glu435 44o Arg Lys Gly Ile Gln Ala Glu Glu Val Leu Gln Lys Tyr Leu Lys 456s Glu Ser Val Ser His Ala Ile Leu Gln Thr Asp Gln Ala Leu465 478u Thr Glu Lys Lys Lys Lys Glu Ala Gln Val Lys Ala Glu Ala 485 49u Lys Ala GluAla Gln Arg Leu Ala Ala Ile Gln Arg Gln Asn Glu 55et Met Gln Glu Arg Glu Arg Leu His Gln Glu Gln Val Arg Gln 5525Met Glu Ile Ala Lys Gln Asn Trp Leu Ala Glu Gln Gln Lys Met Gln 534n Gln Met Gln Glu Gln Ala Ala Gln LeuSer Thr Thr Phe Gln545 556n Asn Arg Ser Leu Leu Ser Glu Leu Gln His Ala Gln Arg Thr 565 57l Asn Asn Asp Asp Pro Cys Val Leu Leu 58828DNAhomo sapiens 63gggggagtga aagcgaaagc ccgggcgact agccgggaga ccagagatct agcgactgaa6tggc caagccgtgt ggggtgcgcc tgagcgggga agcccgcaaa caggtggagg caggca gaatcttttc caggaggctg aggaattcct ctacagattc ttgccacaga catata cctgaatcag ctcttgcaag aggactccct caatgtggct gacttgactt 24gggc cccactggac atccccatcc cagaccctccacccaaggat gatgagatgg 3gataa gcaggagaag aaagaagtcc ctaagtgtgg atttctccct gggaatgaga 36tgtc cctgcttgcc ctggttaagc cagaagtctg gactctcaaa gagaaatgca 42tgat tacatggatc caacacctga tccccaagat tgaagatgga aatgattttg 48caat ccaggagaaggtgctggaga gggtgaatgc cgtcaagacc aaagtggaag 54agac aaccatttcc aagtacttct cagaacgtgg ggatgctgtg gccaaggcct 6gagac tcatgtaatg gattaccggg ccttggtgca tgagcgagat gaggcagcct 66agct cagggccatg gtgctggacc tgagggcctt ctatgctgag ctttatcata72gcag caacctggag aaaattgtca ccccaaaggg tgaagaaaag ccatctatgt 78cccg ggactagaag gaaaataaat gatctatatg ttgtgtgg 82864239PRThomo sapiens 64Met Ala Lys Pro Cys Gly Val Arg Leu Ser Gly Glu Ala Arg Lys Glnlu Val Phe Arg Gln Asn LeuPhe Gln Glu Ala Glu Glu Phe Leu 2Tyr Arg Phe Leu Pro Gln Lys Ile Ile Tyr Leu Asn Gln Leu Leu Gln 35 4 Asp Ser Leu Asn Val Ala Asp Leu Thr Ser Leu Arg Ala Pro Leu 5Asp Ile Pro Ile Pro Asp Pro Pro Pro Lys Asp Asp Glu Met Glu Thr65 7Asp Lys Gln Glu Lys Lys Glu Val Pro Lys Cys Gly Phe Leu Pro Gly 85 9 Glu Lys Val Leu Ser Leu Leu Ala Leu Val Lys Pro Glu Val Trp Leu Lys Glu Lys Cys Ile Leu Val Ile Thr Trp Ile Gln His Leu Pro Lys Ile Glu Asp GlyAsn Asp Phe Gly Val Ala Ile Gln Glu Val Leu Glu Arg Val Asn Ala Val Lys Thr Lys Val Glu Ala Phe Gln Thr Thr Ile Ser Lys Tyr Phe Ser Glu Arg Gly Asp Ala Val Ala Ala Ser Lys Glu Thr His Val Met Asp Tyr Arg AlaLeu Val His Arg Asp Glu Ala Ala Tyr Gly Glu Leu Arg Ala Met Val Leu Asp 2rg Ala Phe Tyr Ala Glu Leu Tyr His Ile Ile Ser Ser Asn Leu 222s Ile Val Thr Pro Lys Gly Glu Glu Lys Pro Ser Met Tyr225 232578DNAhomo sapiens 65cttttgtctc tcagctattt tttgttccct atgtttgtag gatggaaagg cagatgtaaa 6catg gcgaaatata acacgggggg caacccgaca gaggatgtct cagtcaatag cccttc agagtcacag ggccaaactc atcttcagga atacaagcaa gaaagaactt aacaac caaggaaatgccagccctcc tgcaggaccc agcaatgtac ctaagtttgg 24aaag ccacctgtgg cagtcaaacc ttcttctgag gaaaagcctg acaaggaacc 3ccccg tttctaaagc ccactggagc aggccaaaga ttcggaacac cagccagctt 36caga gaccccgagg cgaaagtggg atttctgaaa cctgtaggcc ccaagcccat42gccc aaagaagatt ccaaacctac atttccctgg cctcctggaa acaagccatc 48cagt gtaaaccaag accatgactt aaagccacta ggcccgaaat ctgggcctac 54aacc tcagaaaatg aacagaagca agcgtttccc aaattgactg gggttaaagg 6ttatg tcagcatcac aagatcttga acccaagcccctcttcccca aacccgcctt 66gaag ccgcccctaa gtaccgagaa ctcccatgaa gacgaaagcc ccatgaagaa 72ttca tcaaaagggt ccccagctcc cctgggagtc aggtccaaaa gcggcccttt 78agca agggaagact cagaaaataa agaccatgca ggggagattt caagtttgcc 84tgga gtggttttgaaacctgctgc gagcagggga ggcccaggtg tctccaaaaa 9aagaa aaaaaggaag ataggaagat agatgctgct aagaacacct tccagagcaa 96tcag gaagagttgg cctcagggac tcctcctgcc aggttcccta aggccccttc gctgaca gtgggggggc catggggcca aagtcaggaa aaggaaaagg gagacaagaaagccacc ccgaaacaga agccattgcc tcccttgttt accttgggtc cacctccacc acccaac agaccaccaa atgttgacct gacgaaattc cacaaaacct cttctggaaa tactagc aaaggccaga cgtcttactc aacaacttcc ctgccaccac ctccaccatc tccggcc agccaaccac cattgccagcatctcaccca tcacaaccac cagtcccaag acctccc agaaacatta aacctccgtt tgacctaaaa agccctgtca atgaagacaa agatggt gtcacgcact ctgatggtgc tggaaatcta gatgaggaac aagacagtga agaaaca tatgaagaca tagaagcatc caaagaaaga gagaagaaaa gggaaaaggaaaagaag aggttagagc tggagaaaaa ggaacagaaa gagaaagaaa agaaagaaca aataaag aagaaattta aactaacagg ccctattcaa gtcatccatc ttgcaaaagc ttgtgat gtcaaaggag gaaagaatga actgagcttc aagcaaggag agcaaattga catccgc atcacagaca acccagaaggaaaatggttg ggcagaacag caaggggttc tggctat attaaaacaa ctgctgtaga gattgactat gattctttga aactgaaaaa ctctctt ggtgcccctt caagacctat tgaagatgac caagaagtat atgatgatgt agagcag gatgatatta gcagccacag tcagagtgga agtggaggga tattccctccaccagat gatgacattt atgatgggat tgaagaggaa gatgctgatg atggtttccc tcctcct aaacaattgg acatgggaga tgaagtttac gatgatgtgg atacctctga 2cctgtt tcatcagcag agatgagtca aggaactaat tttggaaaag ctaagacaga 2aaggac cttaagaagc taaaaaagcaggaaaaagaa gaaaaagact tcaggaaaaa 2aaatat gatggtgaaa ttagagtcct atattcaact aaagttacaa cttccataac 222aaag tggggaacca gagatctaca ggtaaaacct ggtgaatctc tagaagttat 228caca gatgacacaa aagttctctg cagaaatgaa gaagggaaat atggttatgt234gagt tacctagcgg acaatgatgg agagatctat gatgatattg ctgatggctg 24atgac aatgactagc actcaacttt ggtcattctg ctgtgttcat taggtgccaa 246gtct ggattttaat tggcatgtta ttgggtatca agaaaattaa tgcacaaaac 252ttat catttgttat gaaatcccaattatctttac aaagtgttta aagtttga 257866783PRThomo sapiens 66Met Ala Lys Tyr Asn Thr Gly Gly Asn Pro Thr Glu Asp Val Ser Valer Arg Pro Phe Arg Val Thr Gly Pro Asn Ser Ser Ser Gly Ile 2Gln Ala Arg Lys Asn Leu Phe Asn Asn Gln Gly Asn AlaSer Pro Pro 35 4 Gly Pro Ser Asn Val Pro Lys Phe Gly Ser Pro Lys Pro Pro Val 5Ala Val Lys Pro Ser Ser Glu Glu Lys Pro Asp Lys Glu Pro Lys Pro65 7Pro Phe Leu Lys Pro Thr Gly Ala Gly Gln Arg Phe Gly Thr Pro Ala 85 9 Leu Thr ThrArg Asp Pro Glu Ala Lys Val Gly Phe Leu Lys Pro Gly Pro Lys Pro Ile Asn Leu Pro Lys Glu Asp Ser Lys Pro Thr Pro Trp Pro Pro Gly Asn Lys Pro Ser Leu His Ser Val Asn Gln His Asp Leu Lys Pro Leu Gly Pro Lys SerGly Pro Thr Pro Pro Thr Ser Glu Asn Glu Gln Lys Gln Ala Phe Pro Lys Leu Thr Gly Val Gly Lys Phe Met Ser Ala Ser Gln Asp Leu Glu Pro Lys Pro Leu Pro Lys Pro Ala Phe Gly Gln Lys Pro Pro Leu Ser Thr Glu Asn 2is Glu Asp Glu Ser Pro Met Lys Asn Val Ser Ser Ser Lys Gly 222o Ala Pro Leu Gly Val Arg Ser Lys Ser Gly Pro Leu Lys Pro225 234g Glu Asp Ser Glu Asn Lys Asp His Ala Gly Glu Ile Ser Ser 245 25u Pro Phe Pro GlyVal Val Leu Lys Pro Ala Ala Ser Arg Gly Gly 267y Val Ser Lys Asn Gly Glu Glu Lys Lys Glu Asp Arg Lys Ile 275 28p Ala Ala Lys Asn Thr Phe Gln Ser Lys Ile Asn Gln Glu Glu Leu 29er Gly Thr Pro Pro Ala Arg Phe Pro Lys AlaPro Ser Lys Leu33hr Val Gly Gly Pro Trp Gly Gln Ser Gln Glu Lys Glu Lys Gly Asp 325 33s Asn Ser Ala Thr Pro Lys

Gln Lys Pro Leu Pro Pro Leu Phe Thr 345y Pro Pro Pro Pro Lys Pro Asn Arg Pro Pro Asn Val Asp Leu 355 36r Lys Phe His Lys Thr Ser Ser Gly Asn Ser Thr Ser Lys Gly Gln 378r Tyr Ser Thr Thr Ser Leu Pro Pro Pro ProPro Ser His Pro385 39er Gln Pro Pro Leu Pro Ala Ser His Pro Ser Gln Pro Pro Val 44er Leu Pro Pro Arg Asn Ile Lys Pro Pro Phe Asp Leu Lys Ser 423l Asn Glu Asp Asn Gln Asp Gly Val Thr His Ser Asp Gly Ala 435 44y Asn Leu Asp Glu Glu Gln Asp Ser Glu Gly Glu Thr Tyr Glu Asp 456u Ala Ser Lys Glu Arg Glu Lys Lys Arg Glu Lys Glu Glu Lys465 478g Leu Glu Leu Glu Lys Lys Glu Gln Lys Glu Lys Glu Lys Lys 485 49u Gln Glu Ile Lys LysLys Phe Lys Leu Thr Gly Pro Ile Gln Val 55is Leu Ala Lys Ala Cys Cys Asp Val Lys Gly Gly Lys Asn Glu 5525Leu Ser Phe Lys Gln Gly Glu Gln Ile Glu Ile Ile Arg Ile Thr Asp 534o Glu Gly Lys Trp Leu Gly Arg Thr Ala Arg GlySer Tyr Gly545 556e Lys Thr Thr Ala Val Glu Ile Asp Tyr Asp Ser Leu Lys Leu 565 57s Lys Asp Ser Leu Gly Ala Pro Ser Arg Pro Ile Glu Asp Asp Gln 589l Tyr Asp Asp Val Ala Glu Gln Asp Asp Ile Ser Ser His Ser 595 6lnSer Gly Ser Gly Gly Ile Phe Pro Pro Pro Pro Asp Asp Asp Ile 662p Gly Ile Glu Glu Glu Asp Ala Asp Asp Gly Phe Pro Ala Pro625 634s Gln Leu Asp Met Gly Asp Glu Val Tyr Asp Asp Val Asp Thr 645 65r Asp Phe Pro Val Ser SerAla Glu Met Ser Gln Gly Thr Asn Phe 667s Ala Lys Thr Glu Glu Lys Asp Leu Lys Lys Leu Lys Lys Gln 675 68u Lys Glu Glu Lys Asp Phe Arg Lys Lys Phe Lys Tyr Asp Gly Glu 69rg Val Leu Tyr Ser Thr Lys Val Thr Thr Ser Ile ThrSer Lys77ys Trp Gly Thr Arg Asp Leu Gln Val Lys Pro Gly Glu Ser Leu Glu 725 73l Ile Gln Thr Thr Asp Asp Thr Lys Val Leu Cys Arg Asn Glu Glu 745s Tyr Gly Tyr Val Leu Arg Ser Tyr Leu Ala Asp Asn Asp Gly 755 76u IleTyr Asp Asp Ile Ala Asp Gly Cys Ile Tyr Asp Asn Asp 778NAhomo sapiens 67gcggccgctc cgcaggcaga gaagccggga gcgtttgagg cggcggcggc acgagcgatg 6atag agttggagcg gtgccagcag caggcgaacg aggtgacgga aattatgcgt acttcg gcaaggtcctggagcgtggt gtgaagctgg ccgaactgca gcagcgttca aactcc tggatatgag ctcaaccttc aacaagacta cacagaacct ggcccagaag 24tggg agaacatccg ttaccggatc tgcgtggggc tggtggtggt tggtgtcctg 3catcc tgattgtgct gctggtcgtc tttctccctc agagcagtga cagcagtagt36cgga cccaggatgc aggcattgcc tcagggcctg ggaactgacc cagctggtcc 42agaa gcccaatggc tgcactggcc gattctggtc tccaaggacc ttggtgtttg 48ctga cccagcccag tgagtgccaa agggcagccc caacatgtgc acccctgcat 54catg cacagacttg cccttgagca ggccgctgtactggccagct gggcaacccc 6agctc ataaaaat 6PRThomo sapiens 68Met Ala Gly Ile Glu Leu Glu Arg Cys Gln Gln Gln Ala Asn Glu Vallu Ile Met Arg Asn Asn Phe Gly Lys Val Leu Glu Arg Gly Val 2Lys Leu Ala Glu Leu Gln Gln Arg Ser AspGln Leu Leu Asp Met Ser 35 4 Thr Phe Asn Lys Thr Thr Gln Asn Leu Ala Gln Lys Lys Cys Trp 5Glu Asn Ile Arg Tyr Arg Ile Cys Val Gly Leu Val Val Val Gly Val65 7Leu Leu Ile Ile Leu Ile Val Leu Leu Val Val Phe Leu Pro Gln Ser 85 9Asp Ser Ser Ser Ala Pro Arg Thr Gln Asp Ala Gly Ile Ala Ser Pro Gly Asn mo sapiens 69gggtttgggg tagaagggag ggagggggca ggacagtgtg gaatctctag ggtgtatggg 6gggg gcacagttag ttctaagtgg gcttttatgc taaaagcctc tggggatatcttgaaa ataaagatag gtgtcccctc cttgctgtca tctagcccag acactctgct ctctgg ctgtctgctc cctgggaagg ctttaggagg accacccagg acaggatgac 24gcca tctgctctgg agctgggtct cagtgcagag ggacagtgac tgtggatggt 3tctct ggtgggaggt gaggatagaa gtgataaagagctaagagga gcttctggga 36gagg aggtcagtct tgcagtggtg aagccaggac ataggagatg gagcagggct 42ggag gagattctga ggaggatgca ggggaaatct tgtctgttaa tgaaatagga 48tggg gtttggggtg gggtggtcat tgccgtttga gctgctgatt ttcatgagtc 54aaaa ctctcgtgtagggttgacaa tgtggggggg tgggggatcc agcttattct 6tttca agtccattct tggggctggt ggggaggcag gagaataccc ctccctaagc 66tgtg tgccgagctt gctttgtgat gttggcaggg gaggggagac ctgggtggtg 72ttcc ctttatcaaa cccttcagtg ggcacaaaat tgagtgcttg attttaggtt78tttt atgaatgtcc aaatctgtgt ttccccctgc cctcccagac tgtgtggcca 84agtg tctggtttgt gttcatctct ccctcatttc tggagcaggg cctgagaccc 9catct cctatgctct gcatccacgc ctcttttgga cattaaaggt tgattgatgc 96gcac tgtttgggtc tctttgggga tgaggggttggcatggtggc agtggtgcca agtgggt gagggtggag agtctccagg gtggaataga atggggactg aagggaagac ccactag atactgattg gcctgtagca gcacttattt gtgcctaggc ttatgcccct gtagagg aaaactaacc agcagcactc acccttaagg gctcctgggt tctgccttcc gtggtgatggccaggtg aatcatattt tgtgtctttg ggatagtaac tgctaccttc gctgccg tcatggacct gcctgagctt tgctgcttca acttttgccc aaagct 5mo sapiens 7agcg gtggagaatc tcaagcatgt gcatttaatt gaggaatagc agaagggcta 6ccaa gaaaagaagt gtgggtatttttgttaagta aaacagccca agtgcttctg tgggtt tctaccaaga tagaggaaaa gggctgaatt ccctctaagt gggacagccg caggat gtgcttccca gcttcactgg ttaatttgac ctgaacctat ttaaagatcc 24cccc tgaagaccta tccgcactca aattctaaca tgaagaaatc tactcgaatg 3tttactttgaatgag ctctattcgg ttgcatgtta tatgtgattt ccttcctccc 36ttcc actgagcgca cccagtctcc cctagtcttc ctctgtgggt gtgatttttg 42ttac aaacaaaacc cttgaagttc ttggcagatg tgtttgtttc tgtttgcatg 48agat accccaggac aagcggggga ttcatttttc agccattcagttgtttcctc 54ccgc agcaaagtga aaatattctt agcactcaga ctgtacttag agtgttttct 6cagtc tgtacagtct gtaggcagaa ggcctcagaa gaaagtcatg gccactcagt 66tgtg ggctttgtaa gtcctggctc tcccgtcaag gttacccaga ggtaaaagct 72gagt ggggccaggt gtgtttggcactccagatag aaggcaaaat gctcagattc 78gtgc acttgtatgc aacctgtcgg tcgataccta gcatttattt ttccctgaca 84gacc tttccctcac ccaccctaag ctcaaagagt ttagcaaaat tctcttttaa 9cagaa tgccagtaag aggttgaccc ctaccatgga acttctggga tgctaaatac 96atgaacaaaataag ttccttatta taagttcctt atactagcag cttcacctaa attttct ctccagcaat attgacttca ctggggaaaa gccaagagtg tgtggtgagt ttgttct cactcgacct ggctaggact ggctaggagc tgttttttgt acatgaggga tgggctt tcctcagtta tctgaatgtt ttacccaagt gccttcctgctattgtagca tagctca gcttccttgt ccacagggtg aaaaaggact aatgcatttt ccatcagttt aactatg ttagcaaaaa cggcctcctg gtagctcaac ctcctgtacg cgtgtgtgtg aatacac acacaaataa acccctctgt ttttctaaga catcttagct ggatattata agcactt tcataaacaactgtaacaaa tcgcaaagga aagagaaaca aaagcattag tgagaca taaacaggca agagaaagtg tattaggaac tgacagctat caaggaagtt tcagtta caaatgctag gaggaaattt tgccaagaag gatggctcat gaaatatttc tacggga agaggcaata agatcctcta agagaatgag aaagtagggg tgtctaaatgaagatgg gtgtgttgca cgtgtgttag aaggatctca gttgagtgaa ggtttgcact acatcta agttaatgta aatatgtagc actctgacag gtctaccgtg ttgctgaatg tatattt ccaaagtttg caagtcttcc tgtattgtac aaagatgctg ctgcttgata tgtatag caatccagat tagtatgttattaaatttta ttttcttacc tgtattttta tttttac ctgtcctcaa aatattacac ccctgttgga attagattta tatttataaa tcagaaa tctttttaag tgtctctttt tacacatagg ttgatttttt tttcttaaga atgatgt attcttgaaa catttgttac tcattccagg aaacaaaaac ccatataata2ccccac tcagagcctg ttagtcacct ctctagaaga tggcatctca ggagaaggaa 2tttgtg gaagaaggaa tcaccttttt cttgctcaag aattatgctg acttcagccc 2cctgga tctggtcact gagaatcatc aagtgtctag atcctccccc caaaataact 222gtag gtgattttga ttttaaaaaattgacaccaa aaccctgcct gcattgtaat 228cgaa aagaattcat gttcacagaa ctcaacgttc aggctaatat ttacagaagg 234atct aaatcctggt agataactcc tgtatgcttt atccaaagga cacccacagt 24agcat agatataacc aaggatgaat tgattccttc aaagaactgg gaggcacgga246attt tttgtttaca tccagtagcc aagacgcctc agtgagccag tcttgggcag 252tcac atttaggcag attggaagtt ggtatgttct aattctcact ctggactaca 258ctga atttatcatg tcaaaaaaaa aaaaaaaaaa agacctttcc aagtgctttc 264tcag aattgaaaga atgttttcatttcaagttta caagaggcat ggatggagtt 27gttct tgacaagctg ggctaacctt tcccgaactt gtttcccgga ggcaaggtgc 276accc agcgcatctt aaccttgggt ctcctaggct cgaggctagg gcattacgtt 282aacc aaagcagcca attgcatagc aagtattttc ctgcattcca attaaatgct288aaag cagcatccta taaaattgtg atcataaaca tccatttccc tcagcttttg 294cctt gacttacagc caacatcact gtttaactca gtctgtttaa aaacaaactt 3ggtggt tgataacaga gagttgctcc ctgagccatc agggtcctgg gagctggaag 3agggtt attaacattc tacctttatgcagctgttgg ctgaccagaa taaactccct 3agttca agctttgaat ggaatggatg caaatgatgt tgtttccatt agagcaggtg 3cagcat tctgattggc ctgagcagac cgaggctatg gctgttggga caagcttagc 324gaca tcttgtcaaa gaacctcact cacccctctg gcctctacag ccctcagagg33aaacc aattctccaa caaacaggtc tctccaacat ggtggtgctg gcaggcttag 336aaaa tcctgactgt taaaggcgtt tgaatacatc acattcctat gcaaatgttt 342tcca gtttaatgta gtttattttt cctatatgta aagtattttt atacggcttg 348gata gtttagcaat aaaacagttg gaagcaa35o sapiens 7tctc acaggactgg agagagaatg cggggcagct gggcagggct cacttccagc 6tcac agtactggga gtaagaggtg acctatttat ttttagaagg gggcagtgat acccag ctcctagctt cattcaaggg aggcaggcgc tttggaagtt tgtaaacacc ttctgagtaagggagg agcacttttt ttccaaaaag gaaagaacgt ctctactggg 24tcct cctgatattc agcattagag tagaaaagaa actattgttt ggccacatta 3ggtta gcaggtgctg cagcctttgc cactgttatt atttttaaag ggcagaaatg 36ggtg aagactttgg accaggcaaa agctgggaag ttatcaattccaaaccagat 42ccca ggctcagcca ctgtattgca gaatcatgga tgaatttcag catatttctt 48atgt ctctttttaa acagcagagc cctggcaagt tttgtctcct ggtctgtagt 54acat tttttacgat cttgggaagt tacattcctg gggttatact cagctatcta 6actgt gtgcattttt gtgtccattgtttaaatgta atgatattgg acaaaaaatt 66aaaa ttaagtcagt tctgctgaaa ctggattttg gaattggaga atatattaat 72aaac gtgagagatc tgaagcagac aaagaaaaaa gtcacaaaga tgacagtgaa 78tttt cagctctttg tcctaagatt agcctcacgg ttgctgccaa agagttatct 84gacacagacgtctc agaggtatcc tggactgata atgggacctt caacctttca 9ataca ctccacagac agacacttct gatgatcttg accgacccag tgaggaagtt 96agag atctttcaga ttttccatct ctagaaaatg gcatgggaac aaatgatgaa gaattaa gccttggttt gcccactgag ctcaagagaa agaaggaacagttggacagt cacagac caagcaaaga gacgcaatca gcagctggtc tcacccttcc tctgaacagt caaacct ttcacctgat gagcaacctg gctggggatg ttatcacagc tgcagtgact gctatca aagaccagtt agagggtgtg cagcaagcac tttctcaggc tgcccccatc gaagagg acacagacactgaagaaggt gatgactttg aactacttga ccagtcagag gatcaaa ttgagagtga attgggactt acacaagacc aggaagcaga agcacagcaa aagaagt cttcaggttt cctttcaaat ctgctgggag gccattaatc taggaatcag gcaacag agcacaaaaa acaccaaaaa aatttcaaac aagaaaaaaa aaaaaaaaagaagaaaa aaattgaact gtaagcttta atgattactt tagatttgtt ttattttccc tgcagtg aattaattgg atatatatca gctgacactg atagattgat atttctgatc atttttg tgtcataagc atggaaatga actttataca caccactgtg ttgtcagaga atattag gggttgtttt taaagcaaaaagaaaaaaac aaaaaccaaa ctattaaaat cctataa atattctttt tctttacagt ttttcaagca tgcaaaacag tttattgtaa actgaaa aatattaaca attaattgtg aatacatgct gttaccagct tccttattcc tacctgg aaaatttttt tttcaacgga tagattttga tgtaaaaaag accgaaattaaggtatc ttagttgaag gacttgggaa atactatcaa aattaatttc ttaggaaaaa taaaagt atatttaagt actctggata gactgaaacg tttccatgtt atttctgcag 2agactt aggcttattt gtaaagaagc atgctccatt gactgccatc tctagtcttg 2gggtgg tattaaccca tagaaagcaagcagttgtgt atcacataga caatggttat 2taaaca gattcagttg ttttgttgtt cattcgtcat atgtttgtga tagggatgtt 222acag ctctattctg cctgctcaga cttaagttag acccttatct tttatattat 228aaaa aagtctccta aaattgtgaa actagttctt gatgagtgat gtgatcatca234aaga tataataact ctgttttctt agcctgtata gaggagagga acttgcttgg 24aaata tatttatttg ccatttaagt ataaatatga aatctgtttc ttattgggaa 246atat atatattttc ctttaaactt tttaaggtca cttttaaata accaaatttg 252ggtt tttaacaaag gactaaagagctgaaaccaa cctagttttg tttttgtgat 258ttta agtgtcgagg gaccatgcca gcaactacca aaaatctctt aaatcttcag 264ctgg cattttggca gatgcataga gacatctgag accctcagaa aggaaggata 27agaat ataggaaatc tgtgttctct tcctttcatt ttatccctta tatttctaaa276ttat aagtaatctg acattttaat gtagctactc ttatttattt tttctttctg 282taaa atatctggac tgagttttgc caaatgttaa agggagaaga gttactgaag 288aaca cttgcttttt gtgattgctt atgtcattag tgcctcatga ctgtgtttga 294ttat tgatacaaag tgagcctgtgccttcattat cttgcccatt ttaatacaaa 3aacctg gtgtttgaaa atctctgaac tgtgtgggtt ttggaggaat atacctgaat 3ttcaat aacagtttct ggacaggaag aaaaatacag ttacatattt ataaaatagt 3atcagt atttttttat gtgtatgttt ctttctttaa aacaatattc ttggatataa3gaaaag tttaaaggtc atttccattt cttcactaag gagaaaaaaa gttaaataat 324aatt aaagaatata agtcactaga tgaccttaca ggaagacgaa ctcaagggct 33tctgt ggtggtatga acaataaatc tagaataaaa tgttaataac tacaaattaa 336tgtg aggatgggag gaagttggtagggtagaaaa atgtgctatt accactattg 342catg ccagctctct agggacagca gcattataac tatgtatgaa ttttaatatt 348atat gactgtatga ctacaaattt acacaataca ataatgggac tttctcacaa 354attc aaacaaacac aaggatgttg aaggttcttg tttgtgtata tgtgtgtttt36tgggg ggtcactgtt tctggtttta aaagatgaag gagcagatac atttcatatg 366ccag tgtagtagag gactacatgt cctttactat gagaatataa atagcaatat 372RThomo sapiens 72Met Pro Glu Gly Glu Asp Phe Gly Pro Gly Lys Ser Trp Glu Val Ileer Lys ProAsp Glu Arg Pro Arg Leu Ser His Cys Ile Ala Glu 2Ser Trp Met Asn Phe Ser Ile Phe Leu Gln Glu Met Ser Leu Phe Lys 35 4 Gln Ser Pro Gly Lys Phe Cys Leu Leu Val Cys Ser Val Cys Thr 5Phe Phe Thr Ile Leu Gly Ser Tyr Ile Pro Gly Val IleLeu Ser Tyr65 7Leu Leu Leu Leu Cys Ala Phe Leu Cys Pro Leu Phe Lys Cys Asn Asp 85 9 Gly Gln Lys Ile Tyr Ser Lys Ile Lys Ser Val Leu Leu Lys Leu Phe Gly Ile Gly Glu Tyr Ile Asn Gln Lys Lys Arg Glu Arg Ser AlaAsp Lys Glu Lys Ser His Lys Asp Asp Ser Glu Leu Asp Phe Ala Leu Cys Pro Lys Ile Ser Leu Thr Val Ala Ala Lys Glu Leu Ser Val Ser Asp Thr Asp Val Ser Glu Val Ser Trp Thr Asp Asn Gly Phe Asn Leu Ser Glu Gly TyrThr Pro Gln Thr Asp Thr Ser Asp Leu Asp Arg Pro Ser Glu Glu Val Phe Ser Arg Asp Leu Ser Asp 2ro Ser Leu Glu Asn Gly Met Gly Thr Asn Asp Glu Asp Glu Leu 222u Gly Leu Pro Thr Glu Leu Lys Arg Lys Lys Glu Gln LeuAsp225 234y His Arg Pro Ser Lys Glu Thr Gln Ser Ala Ala Gly Leu Thr 245 25u Pro Leu Asn Ser Asp Gln Thr Phe His Leu Met Ser Asn Leu Ala 267p Val Ile Thr Ala Ala Val Thr Ala Ala Ile Lys Asp Gln Leu 275 28u Gly ValGln Gln Ala Leu Ser Gln Ala Ala Pro Ile Pro Glu Glu 29hr Asp Thr Glu Glu Gly Asp Asp Phe Glu Leu Leu Asp Gln Ser33lu Leu Asp Gln Ile Glu Ser Glu Leu Gly Leu Thr Gln Asp Gln Glu 325 33a Glu Ala Gln Gln Asn Lys Lys SerSer Gly Phe Leu Ser Asn Leu 345y Gly His 355734omo sapiens 73gtaagtattt aaatacaatt atttttttct ctcaatggta tagcatattc ctatgcttga 6tagg tctactgaaa aaccattgta aatggacgtt acaggtatgc

tgtatttttg tatttt gttgtattaa gtttgatgaa gctaaaatta gggaactctg aacagatttg aaaaaa tgttttaaag gctttaaaac attagggagg cagtctaggg tgataacgaa 24ttaa gtattaaata cacgaagtta catttttgtt catgtttcat tgtccagaaa 3aggaa actattcagttgtgatcaag caggaaaaaa gaaacaccaa cagttgccag 36tgct ttttagctta aaagcatagt gaagatgctt gaggaagact ttgctacctg 42gtag acagacagac tgagagctat cagcatttga aggcccagcc cttgactctg 48attt gaattttttc tttcccatca aatggcatta acaagattgg gcaaagatga54caaa tttctgtgtt ttttgtttgt ttgtttgttt gttttttctt tgggaactga 6gaggc acgaacacta actcttagca tttttctgta gactttttct tctggccctt 66gcca gcaaaacgcc ccttttctga tcattcgtgc gcagagggcc tcccagtaat 72ctct ccatgctaga gagccttctc tttcctctgaggtttgaact gatgttctgt 78acac cctggcatga cagttacgtg tggtcagccc gctccccagg cccgtccctg 84ccag gtgtgggctc taggcaggcc gacaaggtta cacctcccag agcttgtgat 9ttttc tgacagtcaa agtgtgaagg aacccagact tccccgagcc acggtgttca 96ccac aggaatatgcaagacccatc tccaaaagtt tgtctttgat tttttccaag ttagccc cataagcttt gaatcctgta gttacagtgg cataaaggac tgacaaaacc ataagga aaaacctttt ttttctatga attttttttg ttttttaggg gaaagggatt agaatgt catttaatgt actttgcatc atgtctctag aaatatcttt gtccatagtggtggagt ctctctctct ctctctcttt ttgtttgctt ctgttttctt tcttgtcttc ctttctt ttctttttta tttctggtag caggcctcca tagaacaaat ctaaaacaca accatag taatgtaagg agagcttcag tggcacctca aaacccaccc ttcgagatct caaagac agtctcagaa agctgcactgcccaccggct cagctttcat tcaaaaaggc caaggcc aattctgtct tgaagtcaat gcatgtattt actgtttgac agtaaacccg tgccttc tccacgtcca aggctgtgca ttcgtctaat tagcgtcgtg tatgttttcc tattttt tccaataaaa aagcagtggg atgaaaattg ctttgatata tagcaggtaatgaagct attccatagc acttaactgt agtgaatact gtgtcaccaa ttttgaaatc ttaatgt ttaatgcaaa tccattacat ggtgctatta taggctgaca aaatgattta aaatgtg acaacttggg ctcaattcac tctgctttcc aacagtgtaa atgcatagca tttatct gcatgagaac tatgcactaatctatctgaa gaaaaaaact atatcaactt tatctac tttccgttta cttcaatcct tgcctttttg gtcattgtta taatgccagc aggacag aaagaattat aagaaaacca gcataatacc tgatatatta aaatgtagtg gtgaaat ctgtattata ttgctcttct gaagtaagat ttttctacac cggtagcctt2gtctgt cagtcaggac cttctggtat aggtgatgta aaataaccgt acaatattaa 2tgcgat tccataatgc ttagtgaact gtatgaatat tactcaaagt tatgttagtc 2tttccg acttggttct tgtcagctag gtttaaaggt atttcactga gaacgcaaat 222tttt cttgatttcg gctgttttcagtattttgga ggtatacatt tacttaaatt 228tact cgtgttttgt ttttgttttt gttttttgtt ttctttttcc taggggacaa 234gtgt ttgatttcag aaatcagtac ctggcgagat ttttgtctca aaacgactat 24tttca agaactgtgc tgcgaagaca ctctgagaac atttgcaagt caggggcatt246gacc cttgactgat gctatgcgga gactgataca ttttcttaat ggacaatgtt 252aggt acccatgctt gatctgtctt cacaccagac ctcctcatat taaaaggaaa 258aaaa aaaatgtaag aaatcacatg gctatttagt ttcatgcaca gttgcaatat 264caaa aataaaactc tgtacaaactttgggcccga ttcataagaa aaagaagttt 27taaca cgggattttt ttaatatact ttttttggtc taaatttgaa attacttgct 276atta aataaatttc atctcatttt tttccctaaa ccagcaccca tctgcctttt 282caaa gagttacctt tcccagatta gggggatggt atgtggggag cagatagcgg288ttag aaagataagg gggaccaccc acagctggtc gtgagaacag ggagacagtg 294ggtg ggacctcatc tgtgtgcctg gtatcctgag ttttacatgt agatgcattc 3atttga ttcagaaaaa taaactttcc caaaatgtgt ctgaaccaca agagcataca 3aagtgc tacctctaat ctaaccagagcaccttcatg gtggaagaca cccaccaggt 3caatgt gaacttttgt atctctgcag tggtttcaag gacaaatagt gtccaatgta 3gccatt tttcctgctg tttttatact caacttctca aaatgaaaaa agcttttatt 324ttga cttatttgtg ttgttcttat tttttaaatt tttatttttt gataatagtc33gttag cctttttggg tttttttttt ttttttttgg cttttttttt tgtttgtttt 336tttt gacattgcaa ccgaaggtca taaggccgct agctccgctg ggacagaggc 342gaac taacggctcg gtgccttctc cctggtctca gaccatcgtc tctgcactgc 348attt ggtagcctcg ccactgagatactaactaga cctagactag gagctttatc 354tagg aggtccttta ggaagactct caaaggcaaa tccctgatcc cccgccccac 36gccct gccctctcac cagagcaaaa ttcactgggg acttttccca ccacacatgg 366gtcc actcggaata cctctgtttt ccatttcaaa ttgtaggggg aggggatgga372ccag tgatggtaag agatctgtta tgaaacgaaa caccccccgt gttaataact 378gaaa tctgttttta tgagccgggc cccctgtgcc tctagtatac ttgtattgac 384agtt acccttttag ttttactgtg ttctgtgaaa atttgtaatt ggttgagaat 39tgggc gtccattctt attcaactaaatctccacag gttttttgag ctggtgtgga 396taac tcttgtattc aaccattagt gctaccacct tctcacatta caatacaatt 4gaagca agtactgcat ttcctatgca ac 472DNAhomo sapiens 74ccgctttctc cgcgcggtgc ctgcagggct cccagcgagt ggcagcttgg gaggggccgc 6ggtcagactggcac ctgagcggcc accgcgtccc ggccaggcgg gcagaccgac tcctca cctcgcgcgc ggctgacgca ggcagggcgc ccggcccctc ctggggacca gtgccg gctgggggct gtaggcaccg gacggaagca ggcggtgtga ggaccgacga 24catg gcgggggcgg cctgcgagcc ggtggccagg ccgagcctgacctccatctc 3gggag cttcgcagcc tgtggacctg cgactgcgag ctggccctgc tgccgctggc 36gctg cgcctgcagc ccggtgcctt ccagctgagc ggcgaccagc tcgtggtggc 42cggg gagccggcgg cggcgcgggg gggcttcaac gtcttcggtg acggcctcgt 48cgac gggcagctct accgcctcagcagctacatc aagaggtatg tggaactgac 54ctgt gattataaag actacaggga aactatattg agcaaaccaa tgttgttctt 6atgta cagaccaaaa aagacacctc aaaagaaagg acgtacgcgt ttcttgtaaa 66gcac cccaagataa gaagacagat agagcaaggg atggacatgg tcatctcctc 72tggagaaagttacc ggcttcagtc aatgcaatgt tcctctctct ttcagtttga 78agag gcagtgaaga atttcttccc cccaggaaat gaagtggtta atggagaaaa 84cttt gcatatgaat tcaaagctga tgcattattt gatttcttct attggtttgg 9gtaat tccgttgtaa aagtaaatgg aaaagttctg aatttgtcaagtacaagtcc 96gaag gagacgatta agttatttct ggaaaaaatg agtgagcctt taatccgaag cagtttc tctgaccgaa agttcagtgt aacttccaga ggttcaatag atgatgtttt ctgcaat ctgtcaccca gatcatctct gacagagcct cttttggcag aattaccatt aagtgtt ctggaatctgaagagacacc caaccaattt atctgattga actgaacatt gcagttg ctcccgcact ccaggcctgt gctagactat aggctggggg gagggtagga gggaggc agatacttcc acctgcgtgt caatctccgg ctcctccatg gcttctatgg actcctc tcttctgctt ctgtggatgt gatgccctgg caggcccagg gcagctgattctaaaac ttatgattac caggatggaa aggccttggt cccatggcac tgggtggggc gggatat tctctacttt gaacacttct ccaaagaggc agaagggcca cagagttctg ccctgaa catttttctc agttccctgg gagtttttgt ggcagccttt gtgggagtgg gactggc tgttgaccta gcatgcttcataaatcaggg tttggccctc tgcttggagc caacccc ttgaactcaa acctgtcgag caaggggtta agagttctgt tctcttgcca tggctgg gcaaaagcct gtgccatctt tcactgggag gcaaatatgt ttttcatcct atatgac acctatgaga aacgttcaca gtgaggagta gccaggttgc taggacagtactgccac acactgcctg aaatcggaac tcccttggcc tccctcttaa ctaagtgacc gtagaag gaagccagga gatatggtac cgaacaatga caggggaagg gtattggaca cagcgtc ctccttattg aaaacacatt atgtcagttg ggaattttaa ataagctttt aaaccta acactaaaag caaaatagaagaaagctata ccattaccat aatacatttt 2ctcatg gctacaatgg aattcttgaa aaggaaaaaa aaatcctatc tacatataaa 2tgcatg aatgaatcac tacatatgct tataatgagg aagagttatg ggtcctgagt 2tttttt atcctttctt aaaaagtttc tgtattatgc attttgataa cactactgat222tcca cttatatttg aaatgttatg taccacattt gcacaattaa aacttttctt 228caac ctagaattga ttaaatttat gactgaggct tcatgtgagc tttccattgt 234tggg tgttgtattt gccttgtaac ttactgaatt acaataagaa ttgtgggttt 24gccac tttctcaaga agcgccttttgaagaacaag gctatgaagt atttgaagaa 246taaa atttgatact gatctttcag aaaagagaag gggaatgcta cttaataaca 252gtta aacatttatt attacactca ataaaaaatg aagagtatta ac 2572753mo sapiens 75Met Ala Gly Ala Ala Cys Glu Pro Val Ala Arg Pro Ser Leu ThrSerer Ser Gly Glu Leu Arg Ser Leu Trp Thr Cys Asp Cys Glu Leu 2Ala Leu Leu Pro Leu Ala Gln Leu Leu Arg Leu Gln Pro Gly Ala Phe 35 4 Leu Ser Gly Asp Gln Leu Val Val Ala Arg Pro Gly Glu Pro Ala 5Ala Ala Arg Gly Gly PheAsn Val Phe Gly Asp Gly Leu Val Arg Leu65 7Asp Gly Gln Leu Tyr Arg Leu Ser Ser Tyr Ile Lys Arg Tyr Val Glu 85 9 Thr Asn Tyr Cys Asp Tyr Lys Asp Tyr Arg Glu Thr Ile Leu Ser Pro Met Leu Phe Phe Ile Asn Val Gln Thr Lys Lys AspThr Ser Glu Arg Thr Tyr Ala Phe Leu Val Asn Thr Arg His Pro Lys Ile Arg Gln Ile Glu Gln Gly Met Asp Met Val Ile Ser Ser Val Ile Gly Glu Ser Tyr Arg Leu Gln Ser Met Gln Cys Ser Ser Leu Phe Gln AspPhe Gln Glu Ala Val Lys Asn Phe Phe Pro Pro Gly Asn Glu Val Asn Gly Glu Asn Leu Ser Phe Ala Tyr Glu Phe Lys Ala Asp 2eu Phe Asp Phe Phe Tyr Trp Phe Gly Leu Ser Asn Ser Val Val 222l Asn Gly Lys Val Leu Asn LeuSer Ser Thr Ser Pro Glu Lys225 234u Thr Ile Lys Leu Phe Leu Glu Lys Met Ser Glu Pro Leu Ile 245 25g Arg Ser Ser Phe Ser Asp Arg Lys Phe Ser Val Thr Ser Arg Gly 267e Asp Asp Val Phe Asn Cys Asn Leu Ser Pro Arg Ser SerLeu 275 28r Glu Pro Leu Leu Ala Glu Leu Pro Phe Pro Ser Val Leu Glu Ser 29lu Thr Pro Asn Gln Phe Ile36homo sapiens 76caggcggcga ggagagcggt gccttgcagg gatgctgcgg gcgggagcac caaccgggga 6ccgg gcgggagaag tccacaccgggaccaccatc atggcagtgg agtttgacgg attgtg atgggttctg attcccgagt gtctgcaggc gaggcggtgg tgaaccgagt gacaag ctgtccccgc tgcacgagcg catctactgt gcactctctg gttcagctgc 24ccaa gccgtggccg acatggccgc ctaccagctg gagctccatg ggatagaact 3aacctccacttgttt tggctgctgc aaatgtggtg agaaatatca gctataaata 36ggac ttgtctgcac atctcatggt agctggctgg gaccaacgtg aaggaggtca 42tgga accctgggag gaatgctgac tcgacagcct tttgccattg gtggctccgg 48cttt atctatggtt atgtggatgc agcatataag ccaggcatgtctcccgagga 54gcgc ttcaccacag acgctattgc tctggccatg agccgggatg gctcaagcgg 6tcatc tacctggtca ctattacagc tgccggtgtg gaccatcgag tcatcttggg 66actg ccaaaattct atgatgagtg aaccttcccc agacttctct ttcttatttt 72aact ctctaggacc aaaacctggtatggtcattg ggaaatgagt gctcagggag 78ctta ggggaggtgg gtgcttccct cctagatgtc agcatacact ctttcttctt 84cagg tctaaaacat ctttcctaga gaaaacaaaa gggactaaac tagaaatata 9cccta tacatgacag gtgatcacgt actgaatgat tttgaagtag tacaaacaat 96tctcattccgcatc atcatgcggt ccatgatgat gaggccgcaa mo sapiens 77Met Leu Arg Ala Gly Ala Pro Thr Gly Asp Leu Pro Arg Ala Gly Gluis Thr Gly Thr Thr Ile Met Ala Val Glu Phe Asp Gly Gly Val 2Val Met Gly Ser Asp Ser Arg Val Ser AlaGly Glu Ala Val Val Asn 35 4 Val Phe Asp Lys Leu Ser Pro Leu His Glu His Ile Tyr Cys Ala 5Leu Ser Gly Ser Ala Ala Asp Ala Gln Ala Val Ala Asp Met Ala Ala65 7Tyr Gln Leu Glu Leu His Gly Ile Glu Leu Glu Glu Pro Pro Leu Val 85 9Ala Ala Ala Asn Val Val Arg Asn Ile Ser Tyr Lys Tyr Arg Glu Leu Ser Ala His Leu Met Val Ala Gly Trp Asp Gln Arg Glu Gly Gln Val Tyr Gly Thr Leu Gly Gly Met Leu Thr Arg Gln Pro Phe Ile Gly Gly Ser Gly Ser ThrPhe Ile Tyr Gly Tyr Val Asp Ala Ala Tyr Lys Pro Gly Met Ser Pro Glu Glu Cys Arg Arg Phe Thr Thr Ala Ile Ala Leu Ala Met Ser Arg Asp Gly Ser Ser Gly Gly Val Tyr Leu Val Thr Ile Thr Ala Ala Gly Val Asp His ArgVal Ile 2ly Asn Glu Leu Pro Lys Phe Tyr Asp Glu 286mo sapiens 78ccacgcgtcc gcgctgcgcc acatcccacc ggcccttaca ctgtggtgtc cagcagcatc 6catg gggggacttg aaccctgcag caggctcctg ctcctgcctc tcctgctggc agtggt ctccgtcctgtccaggccca ggcccagagc gattgcagtt gctctacggt ccgggc gtgctggcag ggatcgtgat gggagacctg gtgctgacag tgctcattgc 24cgtg tacttcctgg gccggctggt ccctcggggg cgaggggctg cggaggcagc 3ggaaa cagcgtatca ctgagaccga gtcgccttat caggagctcc agggtcagag36tgtc tacagcgacc tcaacacaca gaggccgtat tacaaatgag cccgaatcat 42cagc aacatgatac ctggatccag ccattcctga agcccaccct gcacctcatt 48ccta ccgcgataca gacccacaga gtgccatccc tgagagacca gaccgctccc 54tctc ctaaaataaa catgaagcac aaaaaaaaaaaaaaaaaaaa aaaaaaaaaa 66PRThomo sapiens 79Met Gly Gly Leu Glu Pro Cys Ser Arg Leu Leu Leu Leu Pro Leu Leula Val Ser Gly Leu Arg Pro Val Gln Ala Gln Ala Gln Ser Asp 2Cys Ser Cys Ser Thr Val Ser Pro Gly Val Leu Ala GlyIle Val Met 35 4 Asp Leu Val Leu Thr Val Leu Ile Ala Leu Ala Val Tyr Phe Leu 5Gly Arg Leu Val Pro Arg Gly Arg Gly Ala Ala Glu Ala Ala Thr Arg65 7Lys Gln Arg Ile Thr Glu Thr Glu Ser Pro Tyr Gln Glu Leu Gln Gly 85 9 Arg Ser AspVal Tyr Ser Asp Leu Asn Thr Gln Arg Pro Tyr Tyr Ahomo sapiens 8gcag cgctcgccac ctccccccgg cctgggcagc gctcgcccgg ggagtccagc 6ctgt ggagctgccg ccatggcccc gcggcgggcg cgcggctgcc ggaccctcgg ccggcg ctgctactgctgctgctgct ccggccgccg gcgacgcggg gcatcacgtg cccccc atgtccgtgg aacacgcaga catctgggtc aagagctaca gcttgtactc 24gcgg tacatttgta actctggttt caagcgtaaa gccggcacgt ccagcctgac 3gcgtg ttgaacaagg ccacgaatgt cgcccactgg acaaccccca gtctcaaatg36agac cctgccctgg ttcaccaaag gccagcgcca ccctccacag taacgacggc 42gacc ccacagccag agagcctctc cccttctgga aaagagcccg cagcttcatc 48ctca aacaacacag cggccacaac agcagctatt gtcccgggct cccagctgat 54aaaa tcaccttcca caggaaccac agagataagcagtcatgagt cctcccacgg 6cctct cagacaacag ccaagaactg ggaactcaca gcatccgcct cccaccagcc 66tgtg tatccacagg gccacagcga caccactgtg gctatctcca cgtccactgt 72gtgt gggctgagcg ctgtgtctct cctggcatgc tacctcaagt caaggcaaac 78gctg gccagcgttgaaatggaagc catggaggct ctgccggtga cttgggggac 84caga gatgaagact tggaaaactg ctctcaccac ctatgaaact cggggaaacc 9agcta agtccggagt gaaggagcct ctctgcttta gctaaagacg actgagaaga 96agga agcgggctcc aggagcaagc tcaccaggcc tctcagaagt cccagcaggacacggac tgccgggtcg gcgcctcctg cgcgagggag caggttctcc gcattcccat caccacc tgcctgcctg tcgtgccttg gacccagggc ccagcttccc aggagagacc ggcttct gagcaggatt tttatttcat tacagtgtga gctgcctgga atacatgtgg tgaaata aaaaccctgc cccgaatcttccgtccctca tcctaacttg cagttcacag aaagtga catacccaaa gctctctgtc aattacaagg cttctcctgg cgtgggagac tacaggg aagacaccag cgtttgggct tctaaccacc ctgtctccag ctgctctgca atggaca gggacctggg aaaggtggga gagatgctga gcccagcgaa tcctctccataggattc aggaagaaga aaactcaact cagtgccatt ttacgaatat atgcgtttat tatactt ccttgtctat tatatctata cattatatat tatttgtatt ttgacattgt ttgtata aacaaaataa aacatctatt ttcaatattt ttaaaatgca 67PRThomo sapiens 8a Pro Arg Arg Ala ArgGly Cys Arg Thr Leu Gly Leu Pro Alaeu Leu Leu Leu Leu Leu Arg Pro Pro Ala Thr Arg Gly Ile Thr 2Cys Pro Pro Pro Met Ser Val Glu His Ala Asp Ile Trp Val Lys Ser 35 4 Ser Leu Tyr Ser Arg Glu Arg Tyr Ile Cys Asn Ser Gly Phe Lys 5Arg Lys Ala Gly Thr Ser Ser Leu Thr Glu Cys Val Leu Asn Lys Ala65 7Thr Asn Val Ala His Trp Thr Thr Pro Ser Leu Lys Cys Ile Arg Asp 85 9 Ala Leu Val His Gln Arg Pro Ala Pro Pro Ser Thr Val Thr Thr Gly Val Thr Pro Gln ProGlu Ser Leu Ser Pro Ser Gly Lys Glu Ala Ala Ser Ser Pro Ser Ser Asn Asn Thr Ala Ala Thr Thr Ala Ile Val Pro Gly Ser Gln Leu Met Pro Ser Lys Ser Pro Ser Thr Gly Thr Thr Glu Ile Ser Ser His Glu Ser Ser His GlyThr Pro Ser Thr Thr Ala Lys Asn Trp Glu Leu Thr Ala Ser Ala Ser His Gln

Pro Gly Val Tyr Pro Gln Gly His Ser Asp Thr Thr Val Ala Ile 2hr Ser Thr Val Leu Leu Cys Gly Leu Ser Ala Val Ser Leu Leu 222s Tyr Leu Lys Ser Arg Gln Thr Pro Pro Leu Ala Ser Val Glu225 234u AlaMet Glu Ala Leu Pro Val Thr Trp Gly Thr Ser Ser Arg 245 25p Glu Asp Leu Glu Asn Cys Ser His His Leu 2624DNAArtificial SequenceArtificial Sequence equals primer. 82ggaacaggaa agacttctca agca 248323DNAArtificial SequenceArtificial Sequenceequals primer. 83cttgacgtag ttgcaagctc tca 23842ificial SequenceArtificial Sequence equals primer. 84gctgaagcaa ggtagcgatg a 2AArtificial SequenceArtificial Sequence equals primer. 85cctcgttgct gagtgttgga 2AArtificialSequenceArtificial Sequence equals primer. 86tcagactgaa gagctactgt aatgatca 288726DNAArtificial SequenceArtificial Sequence equals primer. 87caacaatcaa gacattcttt ccagtt 268822DNAArtificial SequenceArtificial Sequence equals primer. 88tgctttcacttgtgcctctt tc 22892ificial SequenceArtificial Sequence equals primer. 89caggctctca cagagacgga a 2AArtificial SequenceArtificial Sequence equals primer. 9gcca catcgct NAArtificial SequenceArtificial Sequence equals primer. 9aggc gcccaatac

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