Poly(ADP-ribosyl)ation affects stabilization of Che-1 protein in response to DNA damage.

Maria Giulia Bacalini (1, 2) , Debora Di Lonardo (2) , Angela Catizone (3) , Fabio Ciccarone (1, 2) , Tiziana Bruno (4) , Michele Zampieri (1, 2) , Tiziana Guastafierro (1, 2) , Roberta Calabrese (1, 2) , Maurizio Fanciulli (4) , Claudio Passananti (5) , Paola Caiafa (1, 2) , Anna Reale (1, 2)

1 Institut Pasteur, Fondation Cenci Bolognetti - Istituto Pasteur Italia, Fondazione Cenci Bolognetti
2 Department of Cellular Biotechnologies and Haematology, Section of Clinical Biochemistry
3 Department of Anatomy, Histology, Forensic Medicine and Orthopedics
4 Laboratory B, Department of Therapeutic Programs Development
5 CNR - National Research Council of Italy | Consiglio Nazionale delle Ricerche

Poly(ADP-ribose) polymerase 1 (PARP-1) catalyzes a post-translational modification that plays a crucial role in coordinating the signalling cascade in response to stress stimuli. During the DNA damage response, phosphorylation by ataxia telangiectasia mutated (ATM) kinase and checkpoint kinase Chk2 induces the stabilization of Che-1 protein, which is critical for the maintenance of G2/M arrest. In this study we showed that poly(ADP-ribosyl)ation, beyond phosphorylation, is involved in the regulation of Che-1 stabilization following DNA damage. We demonstrated that Che-1 accumulation upon doxorubicin treatment is reduced after the inhibition of PARP activity in HCT116 cells and in PARP-1 knock-out or silenced cells. In accordance, impairment in Che-1 accumulation by PARP inhibition reduced Che-1 occupancy at p21 promoter and affected the expression of the corresponding gene. Epistasis experiments showed that the effect of poly(ADP-ribosyl)ation on Che-1 stabilization is independent from ATM kinase activity. Indeed we demonstrated that Che-1 protein co-immunoprecipitates with ADP-ribose polymers and that PARP-1 directly interacts with Che-1, promoting its modification in vitro and in vivo.

Domaines: Biochimie, Biologie Moléculaire

Maria Giulia Bacalini, Debora Di Lonardo, Angela Catizone, Fabio Ciccarone, Tiziana Bruno, et al.. Poly(ADP-ribosyl)ation affects stabilization of Che-1 protein in response to DNA damage.. DNA Repair, 2011, 10 (4), pp.380-9. ⟨10.1016/j.dnarep.2011.01.002⟩. ⟨pasteur-00976870⟩