title: ATR- and ATM-Mediated DNA Damage Response Is Dependent on Excision Repair Assembly during G1 but Not in S Phase of Cell Cycle.
abstract: Cell cycle checkpoint is mediated by ATR and ATM kinases, as a prompt early response to a variety of DNA insults, and culminates in a highly orchestrated signal transduction cascade. Previously, we defined the regulatory role of nucleotide excision repair (NER) factors, DDB2 and XPC, in checkpoint and ATR/ATM-dependent repair pathway via ATR and ATM phosphorylation and recruitment to ultraviolet radiation (UVR)-induced damage sites. Here, we have dissected the molecular mechanisms of DDB2- and XPC- mediated regulation of ATR and ATM recruitment and activation upon UVR exposures. We show that the ATR and ATM activation and accumulation to UVR-induced damage not only depends on DDB2 and XPC, but also on the NER protein XPA, suggesting that the assembly of an active NER complex is essential for ATR and ATM recruitment. ATR and ATM localization and H2AX phosphorylation at the lesion sites occur as early as ten minutes in asynchronous as well as G1 arrested cells, showing that repair and checkpoint-mediated by ATR and ATM starts early upon UV irradiation. Moreover, our results demonstrated that ATR and ATM recruitment and H2AX phosphorylation are dependent on NER proteins in G1 phase, but not in S phase. We reasoned that in G1 the UVR-induced ssDNA gaps or processed ssDNA, and the bound NER complex promote ATR and ATM recruitment. In S phase, when the UV lesions result in stalled replication forks with long single-stranded DNA, ATR and ATM recruitment to these sites is regulated by different sets of proteins. Taken together, these results provide evidence that UVR-induced ATR and ATM recruitment and activation differ in G1 and S phases due to the existence of distinct types of DNA lesions, which promote assembly of different proteins involved in the process of DNA repair and checkpoint activation.
authors.affiliation: Department of Radiology, The Ohio State University, Columbus, Ohio, 43210, United States of America.
authors.forename: Alo
authors.initials: A
authors.lastname: Ray
authors.affiliation: Department of Radiology, The Ohio State University, Columbus, Ohio, 43210, United States of America.
authors.forename: Chessica
authors.initials: C
authors.lastname: Blevins
authors.affiliation: Department of Radiology, The Ohio State University, Columbus, Ohio, 43210, United States of America.
authors.forename: Gulzar
authors.initials: G
authors.lastname: Wani
authors.affiliation: Department of Radiology, Department of Molecular and Cellular Biochemistry, James Cancer Hospital and Solove Research Institute, The Ohio State University, Columbus, Ohio, 43210, United States of America.
authors.forename: Altaf A
authors.initials: AA
authors.lastname: Wani
mesh_terms: D064007:Ataxia Telangiectasia Mutated Proteins / Q000378:metabolism*; D059447:Cell Cycle Checkpoints / Q000528:radiation effects; D002460:Cell Line; D004249:DNA Damage; D004260:DNA Repair / Q000528:radiation effects; D004268:DNA-Binding Proteins / Q000378:metabolism; D016193:G1 Phase / Q000528:radiation effects; D006657:Histones / Q000378:metabolism; D006801:Humans; D008954:Models, Biological; D010766:Phosphorylation / Q000528:radiation effects; D016196:S Phase / Q000528:radiation effects; D013379:Substrate Specificity / Q000528:radiation effects; D014466:Ultraviolet Rays; D051760:Xeroderma Pigmentosum Group A Protein / Q000378:metabolism
chemical_list: C117669:DDB2 protein, human; D004268:DNA-Binding Proteins; C468783:H2AX protein, human; D006657:Histones; D051760:Xeroderma Pigmentosum Group A Protein; C086984:XPC protein, human; C576597:ATR protein, human; D064007:Ataxia Telangiectasia Mutated Proteins