title: The DNA damage-induced decrease of Bcl-2 is secondary to the activation of apoptotic effector caspases.
abstract: Apoptosis induced by DNA-damaging agents or radiation mainly proceeds through death receptor-independent caspase activation. The release of mitochondrial apoptogenic proteins, such as cytochrome c, into the cytoplasm leading to Apaf1-dependent activation of caspase-9 is a key event in this pathway. The permeability of the mitochondrial outer membrane is regulated by the various pro- and antiapoptotic Bcl-2 family proteins, and it is thought that DNA damage triggers apoptosis through the downregulation of antiapoptotic Bcl-2. Using murine embryonic fibroblasts (MEF) deficient and proficient in Apaf1, we show that DNA-damaging agents and radiation lead to a decline in Bcl-2 protein only in wt MEF, but not in apaf1(-/-) MEF, which are defective in the activation of effector caspases and apoptosis. In contrast, the induction of proapoptotic Noxa, the activation of Bax, the cytoplasmic release of cytochrome c, as well as a drop of the mitochondrial transmembrane potential Deltapsim are equally observed in wt and apaf1(-/-) MEF following DNA damage. Moreover, the loss of Bcl-2 protein occurring in wt MEF can be prevented by caspase inhibition. Hence, the activation of proapoptotic Bcl-2 family proteins rather than the downregulation of antiapoptotic Bcl-2 mediates the primary signal in the DNA damage-induced release of mitochondrial apoptogenic proteins in MEF.
authors.affiliation: Gene Therapy Laboratory, Johannes Gutenberg University, 55101 Mainz, Germany.
authors.forename: Javorina
authors.initials: J
authors.lastname: Milosevic
authors.forename: Sandra
authors.initials: S
authors.lastname: Hoffarth
authors.forename: Claudia
authors.initials: C
authors.lastname: Huber
authors.forename: Martin
authors.initials: M
authors.lastname: Schuler
mesh_terms: D000230:Adenocarcinoma / Q000235:genetics / Q000473:pathology; D000256:Adenoviridae / Q000235:genetics; D000590:Amino Acid Chloromethyl Ketones / Q000494:pharmacology; D000818:Animals; D017209:Apoptosis / Q000235:genetics*; D053477:Apoptotic Protease-Activating Factor 1; D061945:Caspase Inhibitors; D020169:Caspases / Q000378:metabolism*; D017690:Cell Transplantation; D004249:DNA Damage; D003609:Dactinomycin / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004307:Dose-Response Relationship, Radiation; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005047:Etoposide / Q000494:pharmacology; D005347:Fibroblasts / Q000187:drug effects / Q000528:radiation effects; D017353:Gene Deletion; D006801:Humans; D051379:Mice; D016513:Mice, SCID; D010190:Pancreatic Neoplasms / Q000235:genetics / Q000473:pathology; D011506:Proteins / Q000187:drug effects / Q000235:genetics / Q000378:metabolism / Q000528:radiation effects; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000187:drug effects* / Q000235:genetics / Q000528:radiation effects*; D014183:Transplantation, Heterologous; D014407:Tumor Cells, Cultured; D014466:Ultraviolet Rays / Q000009:adverse effects
chemical_list: C506592:APAF1 protein, human; D000590:Amino Acid Chloromethyl Ketones; C506593:Apaf1 protein, mouse; D053477:Apoptotic Protease-Activating Factor 1; D061945:Caspase Inhibitors; D004791:Enzyme Inhibitors; D011506:Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; C096713:benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone; D003609:Dactinomycin; D005047:Etoposide; D020169:Caspases