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. 1999 Jan 15;18(2):353–362. doi: 10.1093/emboj/18.2.353

Structure-function analysis of an evolutionary conserved protein, DAP3, which mediates TNF-alpha- and Fas-induced cell death.

J L Kissil 1, O Cohen 1, T Raveh 1, A Kimchi 1
PMCID: PMC1171130  PMID: 9889192

Abstract

A novel approach to the isolation of positive mediators of programmed cell death, based on random inactivation of genes by expression of anti sense RNAs, was employed to identify mediators of interferon-gamma-induced apoptosis. One of the several genes identified is DAP3, which codes for a 46 kDa protein with a potential nucleotide-binding motif. Structure-function studies of the protein indicate that the intact full-length protein is required for its ability to induce apoptosis when overexpressed. The N-terminal 230 amino acids, on the other hand, act in a dominant-negative fashion. Both of these functions are dependent on the integrity of the nucleotide binding motif. Expression of anti-sense DAP3 RNA and of the dominant interfering form of DAP3 both protected cells from apoptosis induced by activation of Fas and tumor necrosis factor alpha (TNF-alpha) receptors. Thus, DAP3 is implicated as a positive mediator of these death-inducing stimuli. It functions downstream of the receptor signaling complex and its death promoting effects depend on caspase activity. In the nematode Caenorhabditis elegans, a potential homolog of DAP3 showing 35% identity and 64% similarity to the human protein was isolated. Overexpression of the nematode DAP3 cDNA in mammalian cells induced cell death, indicating that the protein is conserved at the functional level as well as the structural level.

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Selected References

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  1. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. doi: 10.1016/S0022-2836(05)80360-2. [DOI] [PubMed] [Google Scholar]
  2. Ashkenazi A., Dixit V. M. Death receptors: signaling and modulation. Science. 1998 Aug 28;281(5381):1305–1308. doi: 10.1126/science.281.5381.1305. [DOI] [PubMed] [Google Scholar]
  3. Boldin M. P., Goncharov T. M., Goltsev Y. V., Wallach D. Involvement of MACH, a novel MORT1/FADD-interacting protease, in Fas/APO-1- and TNF receptor-induced cell death. Cell. 1996 Jun 14;85(6):803–815. doi: 10.1016/s0092-8674(00)81265-9. [DOI] [PubMed] [Google Scholar]
  4. Boldin M. P., Varfolomeev E. E., Pancer Z., Mett I. L., Camonis J. H., Wallach D. A novel protein that interacts with the death domain of Fas/APO1 contains a sequence motif related to the death domain. J Biol Chem. 1995 Apr 7;270(14):7795–7798. doi: 10.1074/jbc.270.14.7795. [DOI] [PubMed] [Google Scholar]
  5. Chinnaiyan A. M., O'Rourke K., Tewari M., Dixit V. M. FADD, a novel death domain-containing protein, interacts with the death domain of Fas and initiates apoptosis. Cell. 1995 May 19;81(4):505–512. doi: 10.1016/0092-8674(95)90071-3. [DOI] [PubMed] [Google Scholar]
  6. Cohen O., Feinstein E., Kimchi A. DAP-kinase is a Ca2+/calmodulin-dependent, cytoskeletal-associated protein kinase, with cell death-inducing functions that depend on its catalytic activity. EMBO J. 1997 Mar 3;16(5):998–1008. doi: 10.1093/emboj/16.5.998. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Deiss L. P., Feinstein E., Berissi H., Cohen O., Kimchi A. Identification of a novel serine/threonine kinase and a novel 15-kD protein as potential mediators of the gamma interferon-induced cell death. Genes Dev. 1995 Jan 1;9(1):15–30. doi: 10.1101/gad.9.1.15. [DOI] [PubMed] [Google Scholar]
  8. Deiss L. P., Kimchi A. A genetic tool used to identify thioredoxin as a mediator of a growth inhibitory signal. Science. 1991 Apr 5;252(5002):117–120. doi: 10.1126/science.1901424. [DOI] [PubMed] [Google Scholar]
  9. Finlay C. Rat embryo fibroblast complementation assay with ras genes. Methods Enzymol. 1995;255:389–394. doi: 10.1016/s0076-6879(95)55041-0. [DOI] [PubMed] [Google Scholar]
  10. Gabig T. G., Mantel P. L., Rosli R., Crean C. D. Requiem: a novel zinc finger gene essential for apoptosis in myeloid cells. J Biol Chem. 1994 Nov 25;269(47):29515–29519. [PubMed] [Google Scholar]
  11. Gudkov A. V., Kazarov A. R., Thimmapaya R., Axenovich S. A., Mazo I. A., Roninson I. B. Cloning mammalian genes by expression selection of genetic suppressor elements: association of kinesin with drug resistance and cell immortalization. Proc Natl Acad Sci U S A. 1994 Apr 26;91(9):3744–3748. doi: 10.1073/pnas.91.9.3744. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hardwick J. M. Viral interference with apoptosis. Semin Cell Dev Biol. 1998 Jun;9(3):339–349. doi: 10.1006/scdb.1998.0243. [DOI] [PubMed] [Google Scholar]
  13. Hengartner M. O., Horvitz H. R. Programmed cell death in Caenorhabditis elegans. Curr Opin Genet Dev. 1994 Aug;4(4):581–586. doi: 10.1016/0959-437x(94)90076-f. [DOI] [PubMed] [Google Scholar]
  14. Itoh N., Yonehara S., Ishii A., Yonehara M., Mizushima S., Sameshima M., Hase A., Seto Y., Nagata S. The polypeptide encoded by the cDNA for human cell surface antigen Fas can mediate apoptosis. Cell. 1991 Jul 26;66(2):233–243. doi: 10.1016/0092-8674(91)90614-5. [DOI] [PubMed] [Google Scholar]
  15. Kimchi A. DAP genes: novel apoptotic genes isolated by a functional approach to gene cloning. Biochim Biophys Acta. 1998 Apr 17;1377(2):F13–F33. doi: 10.1016/s0304-419x(98)00002-x. [DOI] [PubMed] [Google Scholar]
  16. Kissil J. L., Deiss L. P., Bayewitch M., Raveh T., Khaspekov G., Kimchi A. Isolation of DAP3, a novel mediator of interferon-gamma-induced cell death. J Biol Chem. 1995 Nov 17;270(46):27932–27936. doi: 10.1074/jbc.270.46.27932. [DOI] [PubMed] [Google Scholar]
  17. Kissil J. L., Kimchi A. Assignment of death associated protein 3 (DAP3) to human chromosome 1q21 by in situ hybridization. Cytogenet Cell Genet. 1997;77(3-4):252–252. doi: 10.1159/000134587. [DOI] [PubMed] [Google Scholar]
  18. Kissil J. L., Kimchi A. Death-associated proteins: from gene identification to the analysis of their apoptotic and tumour suppressive functions. Mol Med Today. 1998 Jun;4(6):268–274. doi: 10.1016/s1357-4310(98)01263-5. [DOI] [PubMed] [Google Scholar]
  19. Levy-Strumpf N., Deiss L. P., Berissi H., Kimchi A. DAP-5, a novel homolog of eukaryotic translation initiation factor 4G isolated as a putative modulator of gamma interferon-induced programmed cell death. Mol Cell Biol. 1997 Mar;17(3):1615–1625. doi: 10.1128/mcb.17.3.1615. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Okkema P. G., Fire A. The Caenorhabditis elegans NK-2 class homeoprotein CEH-22 is involved in combinatorial activation of gene expression in pharyngeal muscle. Development. 1994 Aug;120(8):2175–2186. doi: 10.1242/dev.120.8.2175. [DOI] [PubMed] [Google Scholar]
  21. Peter M. E., Heufelder A. E., Hengartner M. O. Advances in apoptosis research. Proc Natl Acad Sci U S A. 1997 Nov 25;94(24):12736–12737. doi: 10.1073/pnas.94.24.12736. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Saraste M., Sibbald P. R., Wittinghofer A. The P-loop--a common motif in ATP- and GTP-binding proteins. Trends Biochem Sci. 1990 Nov;15(11):430–434. doi: 10.1016/0968-0004(90)90281-f. [DOI] [PubMed] [Google Scholar]
  23. Trauth B. C., Klas C., Peters A. M., Matzku S., Möller P., Falk W., Debatin K. M., Krammer P. H. Monoclonal antibody-mediated tumor regression by induction of apoptosis. Science. 1989 Jul 21;245(4915):301–305. doi: 10.1126/science.2787530. [DOI] [PubMed] [Google Scholar]
  24. Villa P., Kaufmann S. H., Earnshaw W. C. Caspases and caspase inhibitors. Trends Biochem Sci. 1997 Oct;22(10):388–393. doi: 10.1016/s0968-0004(97)01107-9. [DOI] [PubMed] [Google Scholar]
  25. Vito P., Lacanà E., D'Adamio L. Interfering with apoptosis: Ca(2+)-binding protein ALG-2 and Alzheimer's disease gene ALG-3. Science. 1996 Jan 26;271(5248):521–525. doi: 10.1126/science.271.5248.521. [DOI] [PubMed] [Google Scholar]
  26. Wallach D., Kovalenko A. V., Varfolomeev E. E., Boldin M. P. Death-inducing functions of ligands of the tumor necrosis factor family: a Sanhedrin verdict. Curr Opin Immunol. 1998 Jun;10(3):279–288. doi: 10.1016/s0952-7915(98)80166-0. [DOI] [PubMed] [Google Scholar]
  27. Wallach D. Preparations of lymphotoxin induce resistance to their own cytotoxic effect. J Immunol. 1984 May;132(5):2464–2469. [PubMed] [Google Scholar]
  28. Wilson R., Ainscough R., Anderson K., Baynes C., Berks M., Bonfield J., Burton J., Connell M., Copsey T., Cooper J. 2.2 Mb of contiguous nucleotide sequence from chromosome III of C. elegans. Nature. 1994 Mar 3;368(6466):32–38. doi: 10.1038/368032a0. [DOI] [PubMed] [Google Scholar]
  29. Yuan J. Transducing signals of life and death. Curr Opin Cell Biol. 1997 Apr;9(2):247–251. doi: 10.1016/s0955-0674(97)80069-5. [DOI] [PubMed] [Google Scholar]

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