Nitric oxide induced poly(ADP-ribose) polymerase cleavage in RAW 264.7 macrophage apoptosis is blocked by Bcl-2.
Endogenously generated or exogenously supplied nitric oxide causes cleavage of poly(ADP-ribose) polymerase (PARP) and apoptotic cell death in RAW 264.7 macrophages. With the use of NO donors such as S-nitrosoglutathione or spermine-NO we established that PARP digestion occurs in parallel with DNA fragmentation, and is preceded by accumulation of the tumor suppressor gene product p53. PARP cleavage in response to lipopolysaccharide and interferon-gamma treatment is prevented by NG-monomethyl-L-arginine, thus proving a NO requirement. Endogenous NO generation, p53 accumulation, and PARP degradation occurred prior to the detection of significant chromatin condensation. In contrast, in stable Bcl-2 transfected cells, NO-initiated PARP cleavage was almost completely blocked. Our data implicate PARP as a proteolytic substrate during NO-mediated apoptotic cell death in RAW 264.7 macrophages and establish Bcl-2 as an efficient signal terminator in this process.
Messmer
U K
UK
University of Erlangen-Nürnberg, Faculty of Medicine, Department of Medicine IV, Germany.
Reimer
D M
DM
Reed
J C
JC
Brüne
B
B
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D001120:Arginine / Q000031:analogs & derivatives / Q000494:pharmacology; D002460:Cell Line; D002843:Chromatin / Q000378:metabolism; D005978:Glutathione / Q000031:analogs & derivatives / Q000378:metabolism; D007371:Interferon-gamma / Q000494:pharmacology; D008070:Lipopolysaccharides / Q000494:pharmacology; D008264:Macrophages / Q000166:cytology / Q000187:drug effects*; D051379:Mice; D009569:Nitric Oxide / Q000494:pharmacology*; D009603:Nitroso Compounds / Q000378:metabolism; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism*; D011518:Proto-Oncogene Proteins / Q000494:pharmacology*; D019253:Proto-Oncogene Proteins c-bcl-2; D026422:S-Nitrosoglutathione; D013096:Spermine / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism; D019323:omega-N-Methylarginine
D002843:Chromatin; D008070:Lipopolysaccharides; D009603:Nitroso Compounds; D011518:Proto-Oncogene Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D016159:Tumor Suppressor Protein p53; D019323:omega-N-Methylarginine; D013096:Spermine; D009569:Nitric Oxide; D026422:S-Nitrosoglutathione; D007371:Interferon-gamma; D001120:Arginine; D011065:Poly(ADP-ribose) Polymerases; D005978:Glutathione