PARP-1-dependent 3-nitrotyrosine protein modification after DNA damage.
3-nitrotyrosine (NO2-Tyr) is thought to be a specific marker of cell injury during oxidative damage. We have evaluated the role of poly(ADP-ribose)polymerase-1 (PARP-1) in protein nitration after treatment of immortalized fibroblasts parp-1+/+ and parp-1-/- with the alkylating agent 2'-methyl-2'-nitroso-urea (MNU). Both cell lines showed increased iNOS expression following MNU treatment in parallel with a selective induction of tyrosine nitration of different proteins. PARP-1 deficient cells displayed a delayed iNOS accumulation, reduced number of nitrated proteins, and a lower global nitrotyrosine "footprint." We have identified the mitochondrial compartment as the major site of oxidative stress during DNA damage, being MnSOD one of the NO2-Tyr-modified proteins, but not in parp-1-/- cells. These results suggest that NO-derived injury can be modulated by proteins involved in the response to genotoxic damage, such as PARP-1, and may account for the limited oxidative injury in parp-1 knockout mice during carcinogenesis and inflammation.
Siles
E
E
Dpto. Biología Experimental, Universidad de Jaén, Spain.
Martinez-Lara
E
E
Núñez
M I
MI
Muñoz-Gámez
J A
JA
Martín-Oliva
D
D
Valenzuela
M T
MT
Peinado
M A
MA
Ruiz de Almodóvar
J M
JM
Javier Oliver
F
F
D000818:Animals; D004249:DNA Damage; D005347:Fibroblasts / Q000166:cytology; D008070:Lipopolysaccharides / Q000494:pharmacology; D008770:Methylnitrosourea / Q000494:pharmacology; D051379:Mice; D018345:Mice, Knockout; D008870:Microtubules / Q000378:metabolism; D008928:Mitochondria / Q000378:metabolism; D009154:Mutation / Q000235:genetics; D041681:NIH 3T3 Cells; D052247:Nitric Oxide Synthase Type II / Q000378:metabolism; D018384:Oxidative Stress; D000071137:Poly (ADP-Ribose) Polymerase-1; D011065:Poly(ADP-ribose) Polymerases / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D013482:Superoxide Dismutase / Q000378:metabolism; D013997:Time Factors; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000378:metabolism
D008070:Lipopolysaccharides; C002744:3-nitrotyrosine; D014443:Tyrosine; D008770:Methylnitrosourea; D052247:Nitric Oxide Synthase Type II; D013482:Superoxide Dismutase; C499321:Parp1 protein, mouse; D000071137:Poly (ADP-Ribose) Polymerase-1; D011065:Poly(ADP-ribose) Polymerases