title: Sequential activation of poly(ADP-ribose) polymerase 1, calpains, and Bax is essential in apoptosis-inducing factor-mediated programmed necrosis.
abstract: Alkylating DNA damage induces a necrotic type of programmed cell death through the poly(ADP-ribose) polymerases (PARP) and apoptosis-inducing factor (AIF). Following PARP activation, AIF is released from mitochondria and translocates to the nucleus, where it causes chromatin condensation and DNA fragmentation. By employing a large panel of gene knockout cells, we identified and describe here two essential molecular links between PARP and AIF: calpains and Bax. Alkylating DNA damage initiated a p53-independent form of death involving PARP-1 but not PARP-2. Once activated, PARP-1 mediated mitochondrial AIF release and necrosis through a mechanism requiring calpains but not cathepsins or caspases. Importantly, single ablation of the proapoptotic Bcl-2 family member Bax, but not Bak, prevented both AIF release and alkylating DNA damage-induced death. Thus, Bax is indispensable for this type of necrosis. Our data also revealed that Bcl-2 regulates N-methyl-N'-nitro-N'-nitrosoguanidine-induced necrosis. Finally, we established the molecular ordering of PARP-1, calpains, Bax, and AIF activation, and we showed that AIF downregulation confers resistance to alkylating DNA damage-induced necrosis. Our data shed new light on the mechanisms regulating AIF-dependent necrosis and support the notion that, like apoptosis, necrosis could be a highly regulated cell death program.
authors.affiliation: Apoptose et Système Immunitaire, CNRS-URA 1961, Institut Pasteur, 25 Rue du Dr. Roux, 75015 Paris, France.
authors.forename: Rana S
authors.initials: RS
authors.lastname: Moubarak
authors.forename: Victor J
authors.initials: VJ
authors.lastname: Yuste
authors.forename: Cédric
authors.initials: C
authors.lastname: Artus
authors.forename: Aïda
authors.initials: A
authors.lastname: Bouharrour
authors.forename: Peter A
authors.initials: PA
authors.lastname: Greer
authors.forename: Josiane
authors.initials: J
authors.lastname: Menissier-de Murcia
authors.forename: Santos A
authors.initials: SA
authors.lastname: Susin
mesh_terms: D000477:Alkylating Agents / Q000494:pharmacology; D000478:Alkylation / Q000187:drug effects; D000818:Animals; D051033:Apoptosis Inducing Factor / Q000378:metabolism*; D002154:Calpain / Q000378:metabolism*; D020169:Caspases / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism; D003600:Cytosol / Q000187:drug effects / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D004789:Enzyme Activation / Q000187:drug effects; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects / Q000201:enzymology; D008769:Methylnitronitrosoguanidine / Q000494:pharmacology; D051379:Mice; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D008954:Models, Biological; D009336:Necrosis / Q000473:pathology*; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism*; D021381:Protein Transport / Q000187:drug effects; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism; D050998:bcl-2 Homologous Antagonist-Killer Protein / Q000378:metabolism; D051028:bcl-2-Associated X Protein / Q000378:metabolism*
chemical_list: D000477:Alkylating Agents; D051033:Apoptosis Inducing Factor; D016159:Tumor Suppressor Protein p53; D050998:bcl-2 Homologous Antagonist-Killer Protein; D051028:bcl-2-Associated X Protein; D008769:Methylnitronitrosoguanidine; D004247:DNA; D011065:Poly(ADP-ribose) Polymerases; D002154:Calpain; D020169:Caspases