title: Death-associated protein 3 localizes to the mitochondria and is involved in the process of mitochondrial fragmentation during cell death.
abstract: Death-associated protein 3 (DAP3) was previously isolated in our laboratory as a positive mediator of cell death. It is a 46-kDa protein containing a GTP binding domain that was shown to be essential for the induction of cell death. DAP3 functions downstream of the receptor signaling complex, and its death-promoting effects depend on caspase activity. Recent reports have suggested that DAP3 is localized to the mitochondria, but no functional significance of this localization has been reported so far. Here, we study the sub-cellular localization and cellular function of human DAP3 (hDAP3). We found that hDAP3 is localized to the mitochondria and, in contrast to cytochrome c, is not released to the cytoplasm following several cell death signals. Overexpression of hDAP3 induced dramatic changes in the mitochondrial structure involving increased fragmentation of the mitochondria. Both the mitochondrial localization of hDAP3 and its GTP-binding activity were essential for the fragmentation. The punctiform mitochondrial morphology was similar to that observed upon treatment of HeLa cells with staurosporine. In fact, reduction of endogenous hDAP3 protein by RNA interference partially attenuated staurosporine-induced mitochondrial fission. Thus, hDAP3 is a necessary component in the molecular pathway that culminates in fragmented mitochondria, probably reflecting its involvement in the fission process. These results, for the first time, provide a specific functional role for hDAP3 in mitochondrial maintenance.
authors.affiliation: Department of Molecular Genetics, Weizmann Institute of Science, Rehovot 76100, Israel.
authors.forename: Zohar
authors.initials: Z
authors.lastname: Mukamel
authors.forename: Adi
authors.initials: A
authors.lastname: Kimchi
mesh_terms: D048868:Adaptor Proteins, Signal Transducing; D020816:Amino Acid Motifs; D000818:Animals; D017209:Apoptosis; D051017:Apoptosis Regulatory Proteins; D015153:Blotting, Western; D016466:CHO Cells; D016923:Cell Death; D002460:Cell Line; D045744:Cell Line, Tumor; D006224:Cricetinae; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005347:Fibroblasts / Q000378:metabolism; D049452:Green Fluorescent Proteins; D006160:Guanosine Triphosphate / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D008164:Luminescent Proteins / Q000378:metabolism; D051379:Mice; D018613:Microscopy, Confocal; D008928:Mitochondria / Q000378:metabolism*; D009154:Mutation; D041681:NIH 3T3 Cells; D011233:Precipitin Tests; D014176:Protein Biosynthesis; D011506:Proteins / Q000378:metabolism; D020543:Proteome; D034622:RNA Interference; D016601:RNA-Binding Proteins; D051381:Rats; D012269:Ribosomal Proteins; D019311:Staurosporine / Q000494:pharmacology; D013347:Subcellular Fractions / Q000378:metabolism; D014162:Transfection
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D051017:Apoptosis Regulatory Proteins; C097041:DAP3 protein, human; C490762:Dap3 protein, mouse; D004791:Enzyme Inhibitors; D008164:Luminescent Proteins; D011506:Proteins; D020543:Proteome; D016601:RNA-Binding Proteins; D012269:Ribosomal Proteins; D049452:Green Fluorescent Proteins; D006160:Guanosine Triphosphate; D019311:Staurosporine