title: Reconstitution of an ATM-dependent checkpoint that inhibits chromosomal DNA replication following DNA damage.
abstract: Cell cycle checkpoints lead to the inhibition of cell cycle progression following DNA damage. A cell-free system derived from Xenopus eggs has been established that reconstitutes the checkpoint pathway inhibiting DNA replication initiation. DNA containing double-strand breaks inhibits replication initiation in a dose-dependent manner. Upon checkpoint activation, a prereplicative complex is assembled that contains ORC, Cdc6, Cdc7, and MCM proteins but lacks Cdc45. The checkpoint is ATM dependent. Cdk2/CyclinE acts downstream of ATM and is downregulated by Cdk2 phosphorylation on tyrosine 15. Cdk2AF/CyclinE is refractory to checkpoint signaling, and Cdc25A overrides the checkpoint and restores DNA replication. This report provides the description of a DNA damage checkpoint pathway that prevents the onset of S phase independently of the transcriptional function of p53 in a vertebrate organism.
authors.affiliation: Department of Genetics and Development, Columbia University, New York, NY 10032, USA.
authors.forename: V
authors.initials: V
authors.lastname: Costanzo
authors.forename: K
authors.initials: K
authors.lastname: Robertson
authors.forename: C Y
authors.initials: CY
authors.lastname: Ying
authors.forename: E
authors.initials: E
authors.lastname: Kim
authors.forename: E
authors.initials: E
authors.lastname: Avvedimento
authors.forename: M
authors.initials: M
authors.lastname: Gottesman
authors.forename: D
authors.initials: D
authors.lastname: Grieco
authors.forename: J
authors.initials: J
authors.lastname: Gautier
mesh_terms: D000730:Androstadienes / Q000494:pharmacology; D000818:Animals; D064007:Ataxia Telangiectasia Mutated Proteins; D042846:CDC2-CDC28 Kinases; D002110:Caffeine / Q000494:pharmacology; D002352:Carrier Proteins / Q000378:metabolism; D018797:Cell Cycle Proteins; D002474:Cell-Free System; D002875:Chromosomes / Q000235:genetics; D019927:Cyclin E / Q000378:metabolism; D051357:Cyclin-Dependent Kinase 2; D018844:Cyclin-Dependent Kinases / Q000235:genetics / Q000378:metabolism; D004247:DNA / Q000235:genetics; D004249:DNA Damage / Q000502:physiology*; D004261:DNA Replication / Q000187:drug effects / Q000502:physiology*; D004268:DNA-Binding Proteins; D004791:Enzyme Inhibitors / Q000494:pharmacology; D008297:Male; D016296:Mutagenesis / Q000502:physiology; D009687:Nuclear Proteins / Q000378:metabolism; D009865:Oocytes; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000235:genetics* / Q000378:metabolism; D018741:Replication Origin / Q000502:physiology; D016196:S Phase / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins; D015398:Signal Transduction / Q000502:physiology*; D013094:Spermatozoa; D014158:Transcription, Genetic / Q000235:genetics; D025521:Tumor Suppressor Proteins; D014443:Tyrosine / Q000378:metabolism; D000077191:Wortmannin; D014981:Xenopus; D029867:Xenopus Proteins
chemical_list: D000730:Androstadienes; C102209:CDC45 protein, S cerevisiae; D002352:Carrier Proteins; D018797:Cell Cycle Proteins; D019927:Cyclin E; D004268:DNA-Binding Proteins; D004791:Enzyme Inhibitors; D009687:Nuclear Proteins; D010726:Phosphodiesterase Inhibitors; D029701:Saccharomyces cerevisiae Proteins; D025521:Tumor Suppressor Proteins; D029867:Xenopus Proteins; D002110:Caffeine; D014443:Tyrosine; D004247:DNA; D064007:Ataxia Telangiectasia Mutated Proteins; D017346:Protein Serine-Threonine Kinases; D042846:CDC2-CDC28 Kinases; C495891:Cdk2 protein, Xenopus; D051357:Cyclin-Dependent Kinase 2; D018844:Cyclin-Dependent Kinases; D000077191:Wortmannin