Novel molecules of the pyrin/nbs/lrr protein family and uses thereof

Abstract

Novel PYRIN-2, PYRIN-3, PYRIN-5, PYRIN-6, PYRIN-7, PYRIN-8, PYRIN-10, and PYRIN-11 polypeptides, proteins, and nucleic acid molecules are disclosed. In addition to isolated PYRIN-2, PYRIN-3, PYRIN-5, PYRIN-6, PYRIN-7, PYRIN-8, PYRIN-10, and PYRIN-11 proteins, the invention further provides PYRIN-2, PYRIN-3, PYRIN-5, PYRIN-6, PYRIN-7, PYRIN-8, PYRIN-10, and PYRIN-11 fusion proteins, antigenic peptides and anti-PYRIN-2, -PYRIN-3, -PYRIN-5, -PYRIN-6, -PYRIN-7, -PYRIN-8, -PYRIN-10, and -PYRIN-11 antibodies. The invention also provides PYRIN-2, PYRIN-3, PYRIN-5, PYRIN-6, PYRIN-7, PYRIN-8, PYRIN-10, and PYRIN-11 nucleic acid molecules, recombinant expression vectors containing a nucleic acid molecule of the invention, host cells into which the expression vectors have been introduced and non-human transgenic animals in which a PYRIN-2, PYRIN-3, PYRIN-5, PYRIN-6, PYRIN-7, PYRIN-8, PYRIN-10, or PYRIN-11 gene has been introduced or disrupted. Diagnostic, screening and therapeutic methods utilizing compositions of the invention are also provided.

1 . An isolated nucleic acid molecule selected from the group consisting of:
 a) a nucleic acid molecule comprising a nucleic acid sequence that is at least 90% identical to SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:25, or a complement thereof;   b) a nucleic acid molecule comprising a nucleic acid sequence encoding a polypeptide comprising at least 50 contiguous amino acids of the amino acid sequence of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, or SEQ ID NO:24;   c) a nucleic acid molecule comprising a nucleotide sequence encoding a polypeptide with an amino acid sequence which is at least 90% identical to the amino acid sequence of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, or SEQ ID NO:24;   d) a nucleic acid molecule comprising at least 100 contiguous nucleotides of the nucleotide sequence of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:25, or a complement thereof; and   e) a nucleic acid molecule that hybridizes to a nucleic acid molecule consisting of the nucleotide sequence of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:7, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, or SEQ ID NO:25 under conditions of incubation at 45° C. in 6.0X SSC followed by washing in 0.2×SSC/0.1% SDS at 65° C., or a complement thereof.   
     
     
         2 . The isolated nucleic acid molecule of  claim 1 , which is selected from the group consisting of:
 a) a nucleic acid comprising the nucleotide sequence of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:25, or a complement thereof; and   b) a nucleic acid molecule that encodes a polypeptide comprising the amino acid sequence of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, or SEQ ID NO:24.   
     
     
         3 . The isolated nucleic acid molecule of  claim 1 , further comprising vector nucleic acid sequences. 
     
     
         4 . A host cell in culture comprising the nucleic acid molecule of  claim 1 . 
     
     
         5 . The nucleic acid molecule of  claim 1 , further comprising nucleic acid sequences encoding a heterologous polypeptide. 
     
     
         6 . An isolated polypeptide selected from the group consisting of:
 a) a polypeptide encoded by a nucleic acid molecule comprising a nucleic acid sequence that is at least 90% identical to SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, or SEQ ID NO:25;   b) a polypeptide comprising a fragment of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, or SEQ ID NO:24, wherein the fragment comprises at least 50 contiguous amino acids of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, or SEQ ID NO:24;   c) a polypeptide comprising a naturally occurrin g  allelic variant of the amino acid sequence of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, or SEQ ID NO:24, wherein the polypeptide is encoded by a nucleic acid molecule that hybridizes to a nucleic acid molecule consisting of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:7, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, or SEQ ID NO:25 under conditions of incubation at 45° C. in 6.0X SSC followed by washing in 0.2×SSC/0.1% SDS at 65° C.;   d) a polypeptide that is encoded by a nucleic acid molecule comprising at least 100 contiguous nucleotides of the nucleotide sequence of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, or SEQ ID NO:25; and   e) a polypeptide comprising an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, or SEQ ID NO:24.   
     
     
         7 . The isolated polypeptide of  claim 6 , comprising the amino acid sequence of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, or SEQ ID NO:24. 
     
     
         8 . The polypeptide of  claim 6 , further comprising heterologous amino acid sequences. 
     
     
         9 . An antibody that selectively binds to a polypeptide of  claim 6 . 
     
     
         10 . A method for producing a polypeptide selected from the group consisting of:
 a) a polypeptide encoded by a nucleic acid molecule comprising a nucleic acid sequence that is at least 90% identical to SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, or SEQ ID NO:25;   b) a polypeptide comprising a fragment of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, or SEQ ID NO:24, wherein the fragment comprises at least 50 contiguous amino acids of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, or SEQ ID NO:24;   c) a polypeptide comprising a naturally occurring allelic variant of the amino acid sequence of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, or SEQ ID NO:24, wherein the polypeptide is encoded by a nucleic acid molecule that hybridizes to a nucleic acid molecule consisting of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:7, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, or SEQ ID NO:25 under conditions of incubation at 45° C. in 6.0X SSC followed by washing in 0.2×SSC/0.1% SDS at 65° C.;   d) a polypeptide that is encoded by a nucleic acid molecule comprising at least 100 contiguous nucleotides of the nucleotide sequence of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, or SEQ ID NO:25; and   e) a polypeptide comprising an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, or SEQ ED NO:24,   the method comprising culturing the host cell of  claim 4  under conditions in which the polypeptide is expressed.   
     
     
         11 . A method for detecting the presence of a polypeptide in a sample, the method comprising:
 (a) contacting the sample with a compound that selectively binds to the polypeptide of  claim 6 ; and   (b) determining whether the compound binds to a polypeptide in the sample.   
     
     
         12 . The method of  claim 11 , wherein the compound which binds to the polypeptide is an antibody. 
     
     
         13 . A kit comprising a compound which selectively binds to a polypeptide of  claim 6  and instructions for use. 
     
     
         14 . A method for detecting the presence of a nucleic acid molecule of  claim 1  in a sample, the method comprising:
 a) contacting the sample with a nucleic acid probe or primer which selectively hybridizes to the nucleic acid molecule of  claim 1 ; and   b) determining whether the nucleic acid probe or primer binds to a nucleic acid molecule in the sample.   
     
     
         15 . The method of  claim 14 , wherein the sample comprises mRNA molecules and is contacted with a nucleic acid probe. 
     
     
         16 . A kit comprising a compound that selectively hybridizes to a nucleic acid molecule of  claim 1  and instructions for use. 
     
     
         17 . A method for identifying a compound that binds to a polypeptide, the method comprising:
 (a) contacting a cell or a sample comprising the polypeptide of  claim 6  with a test compound; and   (b) determining whether the polypeptide binds to the test compound.   
     
     
         18 . A method for identifying a compound which modulates the activity of a polypeptide of  claim 6 , the method comprising:
 (a) contacting a polypeptide of  claim 6  with a test compound; and   (b) determining the effect of the test compound on the activity of the polypeptide to thereby identify a compound which modulates the activity of the polypeptide.   
     
     
         19 . A method for modulating the activity of a polypeptide, the method comprising contacting the polypeptide of  claim 6  or a cell expressing the polypeptide with a compound that binds to the polypeptide in a sufficient concentration to modulate the activity of the polypeptide. 
     
     
         20 . A method of treating a disorder associated with inappropriate apoptosis or an inflammatory disorder, the method comprising:
 (a) selecting an individual that has a disorder associated with inappropriate apoptosis or an inflammatory disorder; and   (b) modulating the expression or activity of the polypeptide of  claim 6 .