title: Poly(ADP-ribosyl)ation affects stabilization of Che-1 protein in response to DNA damage.
abstract: 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.
authors.affiliation: Department of Cellular Biotechnologies and Haematology, Section of Clinical Biochemistry, Sapienza University of Rome, Viale Regina Elena 324, 00161 Rome, Italy.
authors.forename: Maria Giulia
authors.initials: MG
authors.lastname: Bacalini
authors.forename: Debora
authors.initials: D
authors.lastname: Di Lonardo
authors.forename: Angela
authors.initials: A
authors.lastname: Catizone
authors.forename: Fabio
authors.initials: F
authors.lastname: Ciccarone
authors.forename: Tiziana
authors.initials: T
authors.lastname: Bruno
authors.forename: Michele
authors.initials: M
authors.lastname: Zampieri
authors.forename: Tiziana
authors.initials: T
authors.lastname: Guastafierro
authors.forename: Roberta
authors.initials: R
authors.lastname: Calabrese
authors.forename: Maurizio
authors.initials: M
authors.lastname: Fanciulli
authors.forename: Claudio
authors.initials: C
authors.lastname: Passananti
authors.forename: Paola
authors.initials: P
authors.lastname: Caiafa
authors.forename: Anna
authors.initials: A
authors.lastname: Reale
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D051017:Apoptosis Regulatory Proteins / Q000235:genetics / Q000378:metabolism*; D064007:Ataxia Telangiectasia Mutated Proteins; D018797:Cell Cycle Proteins / Q000378:metabolism; D002460:Cell Line; D004249:DNA Damage / Q000187:drug effects; D004268:DNA-Binding Proteins / Q000378:metabolism; D004789:Enzyme Activation / Q000187:drug effects; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D051379:Mice; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D055550:Protein Stability; D016283:Proto-Oncogene Proteins p21(ras) / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D025521:Tumor Suppressor Proteins / Q000378:metabolism
chemical_list: D000970:Antineoplastic Agents; D051017:Apoptosis Regulatory Proteins; D018797:Cell Cycle Proteins; D004268:DNA-Binding Proteins; D011993:Recombinant Fusion Proteins; D025521:Tumor Suppressor Proteins; D011065:Poly(ADP-ribose) Polymerases; C576324:ATM protein, human; D064007:Ataxia Telangiectasia Mutated Proteins; C576325:Atm protein, mouse; D017346:Protein Serine-Threonine Kinases; D016283:Proto-Oncogene Proteins p21(ras)