Feedback-regulated poly(ADP-ribosyl)ation by PARP-1 is required for rapid response to DNA damage in living cells

• Published in: Nucleic acids research 2007-12, Vol.35 (22), p.7665-7675
• Main Authors: Mortusewicz, Oliver, Amé, Jean-Christophe, Schreiber, Valérie, Leonhardt, Heinrich
• Format: Article
• Language: English
• Subjects: Animals; Biochemistry, Molecular Biology; Cell Nucleolus - enzymology; Cells, Cultured; DNA Damage; DNA Repair; DNA-Binding Proteins - metabolism; Feedback, Physiological; Gene Deletion; HeLa Cells; Humans; Kinetics; Life Sciences; Mice; Molecular Biology; Poly (ADP-Ribose) Polymerase-1; Poly Adenosine Diphosphate Ribose - metabolism; Poly(ADP-ribose) Polymerases - chemistry; Poly(ADP-ribose) Polymerases - genetics; Poly(ADP-ribose) Polymerases - metabolism; Protein Structure, Tertiary; X-ray Repair Cross Complementing Protein 1
• ISSN: 0305-1048; 1362-4962
• DOI: 10.1093/nar/gkm933

Genome integrity is constantly threatened by DNA lesions arising from numerous exogenous and endogenous sources. Survival depends on immediate recognition of these lesions and rapid recruitment of repair factors. Using laser microirradiation and live cell microscopy we found that the DNA-damage dependent poly(ADP-ribose) polymerases (PARP) PARP-1 and PARP-2 are recruited to DNA damage sites, however, with different kinetics and roles. With specific PARP inhibitors and mutations, we could show that the initial recruitment of PARP-1 is mediated by the DNA-binding domain. PARP-1 activation and localized poly(ADP-ribose) synthesis then generates binding sites for a second wave of PARP-1 recruitment and for the rapid accumulation of the loading platform XRCC1 at repair sites. Further PARP-1 poly(ADP-ribosyl)ation eventually initiates the release of PARP-1. We conclude that feedback regulated recruitment of PARP-1 and concomitant local poly(ADP-ribosyl)ation at DNA lesions amplifies a signal for rapid recruitment of repair factors enabling efficient restoration of genome integrity.