title: A requirement for replication in activation of the ATR-dependent DNA damage checkpoint.
abstract: Using the Xenopus egg extract system, we investigated the involvement of DNA replication in activation of the DNA damage checkpoint. We show here that DNA damage slows replication in a checkpoint-independent manner and is accompanied by replication-dependent recruitment of ATR and Rad1 to chromatin. We also find that the replication proteins RPA and Polalpha accumulate on chromatin following DNA damage. Finally, damage-induced Chk1 phosphorylation and checkpoint arrest are abrogated when replication is inhibited. These data indicate that replication is required for activation of the DNA damage checkpoint and suggest a unifying model for ATR activation by diverse lesions during S phase.
authors.affiliation: Department of Molecular Pharmacology, Stanford University, Stanford, California 94305-5174, USA.
authors.forename: Patrick J
authors.initials: PJ
authors.lastname: Lupardus
authors.forename: Tony
authors.initials: T
authors.lastname: Byun
authors.forename: Muh-Ching
authors.initials: MC
authors.lastname: Yee
authors.forename: Mohammad
authors.initials: M
authors.lastname: Hekmat-Nejad
authors.forename: Karlene A
authors.initials: KA
authors.lastname: Cimprich
mesh_terms: D000818:Animals; D064007:Ataxia Telangiectasia Mutated Proteins; D002453:Cell Cycle; D018797:Cell Cycle Proteins / Q000378:metabolism*; D000071877:Checkpoint Kinase 1; D004249:DNA Damage; D004261:DNA Replication / Q000528:radiation effects; D004268:DNA-Binding Proteins; D004720:Endonucleases / Q000235:genetics / Q000378:metabolism; D005260:Female; D066298:In Vitro Techniques; D008741:Methyl Methanesulfonate / Q000633:toxicity; D008954:Models, Biological; D009865:Oocytes / Q000187:drug effects / Q000378:metabolism / Q000528:radiation effects; D011494:Protein Kinases / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014466:Ultraviolet Rays / Q000009:adverse effects; D014981:Xenopus; D029867:Xenopus Proteins
chemical_list: D018797:Cell Cycle Proteins; D004268:DNA-Binding Proteins; D011994:Recombinant Proteins; D029867:Xenopus Proteins; D008741:Methyl Methanesulfonate; D011494:Protein Kinases; C576898:Atr protein, Xenopus; D064007:Ataxia Telangiectasia Mutated Proteins; D000071877:Checkpoint Kinase 1; C000605875:Chek1 protein, Xenopus; D017346:Protein Serine-Threonine Kinases; D004720:Endonucleases