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

• Published in: DNA repair 2011-04, Vol.10 (4), p.380-389
• Main Authors: Bacalini, Maria Giulia, Di Lonardo, Debora, Catizone, Angela, Ciccarone, Fabio, Bruno, Tiziana, Zampieri, Michele, Guastafierro, Tiziana, Calabrese, Roberta, Fanciulli, Maurizio, Passananti, Claudio, Caiafa, Paola, Reale, Anna
• Format: Article
• Language: English
• Subjects: Animals; Antineoplastic Agents; Antineoplastic Agents - pharmacology; Apoptosis Regulatory Proteins; Apoptosis Regulatory Proteins - genetics; Apoptosis Regulatory Proteins - metabolism; Ataxia Telangiectasia Mutated Proteins; Bacteriology; Biochemistry, Molecular Biology; Biological and medical sciences; Cell Cycle Proteins; Cell Cycle Proteins - metabolism; Cell Line; Che-1; DNA Damage; DNA Damage - drug effects; DNA damage response; DNA-Binding Proteins; DNA-Binding Proteins - metabolism; Enzyme Activation; Enzyme Activation - drug effects; Fundamental and applied biological sciences. Psychology; Gene Expression Regulation, Neoplastic; Gene Expression Regulation, Neoplastic - drug effects; Growth, nutrition, cell differenciation; Humans; Life Sciences; Mice; Microbiology; Molecular and cellular biology; Molecular genetics; Mutagenesis. Repair; PARP inhibitor; PARP-1; Poly(ADP-ribose) Polymerases; Poly(ADP-ribose) Polymerases - metabolism; Post-translational modification; Promoter Regions, Genetic; Protein Binding; Protein Stability; Protein-Serine-Threonine Kinases; Protein-Serine-Threonine Kinases - metabolism; Proto-Oncogene Proteins p21(ras); Proto-Oncogene Proteins p21(ras) - genetics; Recombinant Fusion Proteins; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - metabolism; Tumor Suppressor Proteins; Tumor Suppressor Proteins - metabolism
• ISSN: 1568-7864; 1568-7856
• DOI: 10.1016/j.dnarep.2011.01.002

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.