Claims
- 1. An isolated polypeptide capable of stimulating NF-kB-dependent transcription or p38-dependent transcription, the polypeptide comprising an amino acid sequence selected from the group consisting of:
(a) an amino acid sequence selected from the group consisting of SEQ ID NO:4, SEQ ID NO:8, and SEQ ID NO:12; (b) an amino acid sequence selected from the group consisting of: amino acids x1 to x2 of SEQ ID NO:4, wherein x1 is any of amino acids 130 through 134 of SEQ ID NO:4, and x2 is any of amino acids 187 through 191 of SEQ ID NO:4; amino acids x1 to x2 of SEQ ID NO:8, wherein x1 is any of the amino acids 132 through 136 of SEQ ID NO:8, and x2 is any of amino acids 189 through 193 of SEQ ID NO:8; and amino acids x1 to x2 of SEQ ID NO:12, wherein x1 is any of the amino acids 155 through 160 of SEQ ID NO:12, and x2 is any of amino acids 212 through 217 of SEQ ID NO:12; (c) an amino acid sequence selected from the group consisting of: amino acids x1 to x2 of SEQ ID NO:4, wherein x1 is any of amino acids 1 through 10 of SEQ ID NO:4, and x2 is any of amino acids 409 through 418 of SEQ ID NO:4; amino acids x1 to x2 of SEQ ID NO:8, wherein x1 is any of amino acids 1 through 10 of SEQ ID NO:8, and x2 is any of amino acids 410 through 419 of SEQ ID NO:8; and amino acids x1 to x2 of SEQ ID NO:12, wherein x1 is any of amino acids 1 through 10 of SEQ ID NO:12, and x2 is any of amino acids 435 through 445 of SEQ ID NO:12; (d) an allelic variant of any of (a)-(c) above; (e) a fragment of the amino acid sequences of any of (a)-(d) comprising at least 20 contiguous amino acids; (f) a fragment of the amino acid sequences of any of (a)-(d), wherein a polypeptide consisting of said fragment is capable of stimulating NF-kB-dependent transcription; (g) a fragment of the amino acid sequences of any of (a)-(d) comprising RING-finger-like domain amino acid sequences; (h) an amino acid sequence comprising at least 20 amino acids and sharing a certain percent amino acid identity with the amino acid sequences of any of (a)-(g), wherein the percent amino acid identity is selected from the group consisting of: at least 85%, at least 90%, at least 95%, at least 97.5%, at least 99%, and at least 99.5%; (i) an amino acid sequence of (h), wherein a polypeptide comprising said amino acid sequence of (h) binds to an antibody that also binds to a polypeptide comprising an amino acid sequence of any of (a)-(g); and (j) an amino acid sequence of (h) or (i) capable of stimulating NF-kB-dependent transcription.
- 2. An isolated nucleic acid encoding a polypeptide of claim 1.
- 3. The nucleic acid of claim 2 comprising a nucleotide sequence selected from the group consisting of:
(a) SEQ ID NO:3; (b) SEQ ID NO:7; (c) SEQ ID NO:11; (d) an allelic variant of (a)-(c); (e) a nucleic acid, having a length of at least 15 nucleotides, that hybridizes under conditions of moderate stringency to the nucleic acid of any of claims (a) through (d); (f) a nucleic acid comprising a nucleotide sequence that shares a certain percent nucleotide sequence identity with the nucleotide sequences of the nucleic acids of any of (a)-(e), wherein the percent nucleotide sequence identity is selected from the group consisting of: at least 85%, at least 90%, at least 95%, at least 97.5%, at least 99%, and at least 99.5%.
- 4. An isolated polypeptide capable of stimulating NF-kB-dependent transcription comprising an amino acid sequence selected from the group consisting of:
(a) an amino acid sequence selected from the group consisting of SEQ ID NO:4, SEQ ID NO:8, and SEQ ID NO:12; (b) an amino acid sequence selected from the group consisting of: amino acids x1 to x2 of SEQ ID NO:4, wherein x1 is any of amino acids 130 through 134 of SEQ ID NO:4, and x2 is any of amino acids 187 through 191 of SEQ ID NO:4; amino acids x1 to x2 of SEQ ID NO:8, wherein x1 is any of the amino acids 132 through 136 of SEQ ID NO:8, and x2 is any of amino acids 189 through 193 of SEQ ID NO:8; and amino acids x1 to x2 of SEQ ID NO:12, wherein x1 is any of the amino acids 155 through 160 of SEQ ID NO:12, and x2 is any of amino acids 212 through 217 of SEQ ID NO:12; (c) an amino acid sequence selected from the group consisting of: amino acids x1 to x2 of SEQ ID NO:4, wherein x1 is any of amino acids 1 through 10 of SEQ ID NO:4, and x2 is any of amino acids 409 through 418 of SEQ ID NO:4; amino acids x1 to x2 of SEQ ID NO:8, wherein x1 is any of amino acids 1 through 10 of SEQ ID NO:8, and x2 is any of amino acids 410 through 419 of SEQ ID NO:8; and amino acids x1 to x2 of SEQ ID NO:12, wherein x1 is any of amino acids 1 through 10 of SEQ ID NO:12, and x2 is any of amino acids 435 through 445 of SEQ ID NO:12; (d) an allelic variant of any of (a)-(c) above; (e) a fragment of the amino acid sequences of any of (a)-(d) comprising at least 20 contiguous amino acids; (f) a fragment of the amino acid sequences of any of (a)-(d), wherein a polypeptide consisting of said fragment is capable of stimulating NF-kB-dependent transcription; and (g) a fragment of the amino acid sequences of any of (a)-(d) comprising RING-finger-like domain amino acid sequences.
- 5. An expression vector comprising at least one nucleic acid according to claim 2.
- 6. A recombinant host cell comprising at least one recombinant nucleic acid comprising the nucleic acid according to claim 2.
- 7. The recombinant host cell of claim 6, wherein the recombinant nucleic acid is integrated into the host cell genome.
- 8. A process for producing a polypeptide encoded by the nucleic acid of claim 2, comprising culturing a recombinant host cell under conditions promoting expression of said polypeptide, wherein the recombinant host cell comprises at least one recombinant nucleic acid comprising the nucleic acid according to claim 2.
- 9. The process of claim 8 further comprising purifying said polypeptide.
- 10. The polypeptide produced by the process of claim 8.
- 11. An isolated antibody that binds to the polypeptide of claim 10.
- 12. An isolated antibody that inhibits the activity of the polypeptide of claim 10.
- 13. An isolated polypeptide capable of inhibiting NF-kB-dependent or p38-dependent transcription, the polypeptide comprising an amino acid sequence selected from the group consisting of:
(a) an amino acid sequence selected from the group consisting of SEQ ID NO:4, SEQ ID NO:8, and SEQ ID NO:12, wherein amino acids 1 through x1 have been deleted from said sequence, and wherein x1 is any of amino acids 50 though 98 of said sequence; (b) SEQ ID NO:4, wherein amino acids x1 through x2 have been deleted from said sequence, and wherein x1 is any amino acid from 99 through 178 and x2 is any amino acid from 100 through 179; (c) SEQ ID NO:8, wherein amino acids x1 through x2 have been deleted from said sequence, and wherein x1 is any amino acid from 1 through 180 and x2 is any amino acid from 2 through 181; (d) SEQ ID NO:12, wherein amino acids x1 through x2 have been deleted from said sequence, and wherein x1 is any amino acid from 1 through 206 and x2 is any amino acid from 2 through 207; (e) an amino acid sequence selected from the group consisting of SEQ ID NO:4, SEQ ID NO:8, and SEQ ID NO:12, wherein one or more cysteine residues of the RING-finger-like domain have been deleted or replaced by non-cysteine residues; (f) an allelic variant of (a)-(e); (g) fragments of the amino acid sequences of any of (a)-(d) and (f) comprising RING-finger-like domain amino acid sequences; and (h) a fragment of the amino acid sequences of any of (a)-(g), wherein a polypeptide consisting of said fragment is capable of inhibiting NF-kB -dependent transcription.
- 14. An isolated nucleic acid encoding a polypeptide of claim 13.
- 15. The nucleic acid of claim 14 comprising a nucleotide sequence selected from the group consisting of
(a) an allelic variant of the nucleic acid of claim 14; (b) a nucleic acid, having a length of at least 15 nucleotides, that hybridizes under conditions of moderate stringency to the nucleic acid of (a); (c) a nucleic acid comprising a nucleotide sequence that shares a certain percent nucleotide sequence identity with the nucleotide sequences of the nucleic acids of any of (a)-(b), wherein the percent nucleotide sequence identity is selected from the group consisting of: at least 85%, at least 90%, at least 95%, at least 97.5%, at least 99%, and at least 99.5%.
- 16. An expression vector comprising at least one nucleic acid according to claim 14.
- 17. A recombinant host cell comprising at least one recombinant nucleic acid comprising the nucleic acid according to claim 14.
- 18. The recombinant host cell of claim 17, wherein the nucleic acid is integrated into the host cell genome.
- 19. A process for producing a polypeptide encoded by the nucleic acid of claim 14, comprising culturing a recombinant host cell under conditions promoting expression of said polypeptide, wherein the recombinant host cell comprises at least one recombinant nucleic acid comprising the nucleic acid according to claim 14.
- 20. The process of claim 19 further comprising purifying said polypeptide.
- 21. The polypeptide produced by the process of claim 19.
- 22. An isolated antibody that binds to the polypeptide of claim 21.
- 23. An isolated antibody that inhibits the activity of the polypeptide of claim 21.
- 24. An isolated nucleic acid encoding the polypeptide of claim 4.
- 25. An isolated genomic nucleic acid corresponding to the nucleic acid of claim 24.
- 26. A method for identifying compounds that alter Pellino polypeptide activity comprising
(a) mixing a test compound with the polypeptide of claim 10; and (b) determining whether the test compound alters the Pellino polypeptide activity of said polypeptide.
- 27. A method for identifying compounds that alter Pellino dominant-negative activity comprising
(a) mixing a test compound with the polypeptide of claim 21; and (b) determining whether the test compound alters the Pellino dominant-negative activity of said polypeptide.
- 28. A method for identifying compounds that alter Pellino polypeptide activity comprising
(a) mixing a test compound with a cell expressing the polypeptide of claim 10; and (b) determining whether the test compound alters the Pellino polypeptide activity of said polypeptide.
- 29. The method of claim 28 further comprising treating the cell with molecule selected from the group consisting of interleukin-1 (IL-1 ), TNF-alpha, IL-18, and phorbol 12-myristate 13-acetate (PMA), peptidoglycan, bacterial lipopeptides, bacterial lipopolysaccharides, zymosan, CpG DNA, flagellin, lipoteichoic acids, and Respiratory Syncytial Virus proteins.
- 30. The method of claim 28 wherein determining whether the test compound alters the Pellino polypeptide activity of said polypeptide comprises determining the level of NF-kB-dependent transcription.
- 31. A method for identifying compounds that alter Pellino dominant-negative activity comprising
(a) mixing a test compound with a cell expressing the polypeptide of claim 21; and (b) determining whether the test compound alters the Pellino dominant-negative activity of said polypeptide.
- 32. The method of claim 31 further comprising treating the cell with molecule selected from the group consisting of interleukin-1 (IL-1 ), TNF-alpha, IL-18, and phorbol 12-myristate 13-acetate (PMA), peptidoglycan, bacterial lipopeptides, bacterial lipopolysaccharides, zymosan, CpG DNA, flagellin, lipoteichoic acids, and Respiratory Syncytial Virus proteins.
- 33. The method of claim 31 wherein determining whether the test compound alters the Pellino dominant-negative activity of said polypeptide comprises determining the level of NF-kB-dependent transcription.
- 34. An isolated polypeptide comprising an amino acid sequence selected from the group consisting of: SEQ ID NO:4, SEQ ID NO:8, and SEQ ID NO:12.
- 35. An isolated nucleic acid encoding the polypeptide of claim 34.
- 36. An isolated polypeptide comprising an amino acid sequence selected from the group consisting of:
(a) an amino acid sequence selected from the group consisting of SEQ ID NO:4, SEQ ID NO:8, and SEQ ID NO:12, wherein amino acids 1 through x1 have been deleted from said sequence, and wherein x1 is any of amino acids 50 though 98 of said sequence; (b) SEQ ID NO:4, wherein amino acids x1 through x2 have been deleted from said sequence, and wherein x1 is any amino acid from 99 through 157 and x2 is any amino acid from 100 through 158; (c) SEQ ID NO:8, wherein amino acids x1 through x2 have been deleted from said sequence, and wherein x1 is any amino acid from 1 through 159 and x2 is any amino acid from 2 through 160; and (d) SEQ ID NO:12, wherein amino acids x1 through x2 have been deleted from said sequence, and wherein x1 is any amino acid from 1 through 184 and x2 is any amino acid from 2 through 185.
- 37. An isolated nucleic acid encoding the polypeptide of claim 36.
Parent Case Info
[0001] This application claims the benefit under 35 U.S.C. 119(e) of U.S. provisional application Serial No. 60/200,198, filed Apr. 28, 2000, which is incorporated by reference herein.
Provisional Applications (1)
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Number |
Date |
Country |
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60200198 |
Apr 2000 |
US |