title: Activation of the DNA replication checkpoint through RNA synthesis by primase.
abstract: When DNA replication is inhibited during the synthesis (S) phase of the cell cycle, a signaling pathway (checkpoint) is activated that serves to prevent mitosis from initiating before completion of replication. This replication checkpoint acts by down-regulating the activity of the mitotic inducer cdc2-cyclin B. Here, we report the relation between chromatin structure and induction of the replication checkpoint. Chromatin was competent to initiate a checkpoint response only after the DNA was unwound and DNA polymerase alpha had been loaded. Checkpoint induction did not require new DNA synthesis on the unwound template strand but did require RNA primer synthesis by primase. These findings identify the RNA portion of the primer as an important component of the signal that activates the replication checkpoint.
authors.affiliation: Department of Biology, University of California, San Diego, La Jolla, CA 92093-0349, USA. matt@mcb.harvard.edu
authors.forename: W M
authors.initials: WM
authors.lastname: Michael
authors.forename: R
authors.initials: R
authors.lastname: Ott
authors.forename: E
authors.initials: E
authors.lastname: Fanning
authors.forename: J
authors.initials: J
authors.lastname: Newport
mesh_terms: D000818:Animals; D016590:Aphidicolin / Q000494:pharmacology; D042846:CDC2-CDC28 Kinases; D002352:Carrier Proteins / Q000378:metabolism; D000071877:Checkpoint Kinase 1; D002843:Chromatin / Q000378:metabolism*; D019927:Cyclin E / Q000378:metabolism; D051357:Cyclin-Dependent Kinase 2; D018844:Cyclin-Dependent Kinases / Q000378:metabolism; D004265:DNA Helicases / Q000378:metabolism; D004256:DNA Polymerase I / Q000037:antagonists & inhibitors / Q000378:metabolism; D019915:DNA Primase / Q000378:metabolism*; D004261:DNA Replication / Q000187:drug effects; D004277:DNA, Single-Stranded / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism; D008938:Mitosis; D009687:Nuclear Proteins / Q000378:metabolism; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D012313:RNA / Q000096:biosynthesis*; D011994:Recombinant Proteins / Q000378:metabolism; D016196:S Phase; D029701:Saccharomyces cerevisiae Proteins; D015398:Signal Transduction; D014981:Xenopus; D029867:Xenopus Proteins
chemical_list: C102209:CDC45 protein, S cerevisiae; D002352:Carrier Proteins; D002843:Chromatin; D019927:Cyclin E; D004277:DNA, Single-Stranded; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; C034477:RNA primers; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; D029867:Xenopus Proteins; D016590:Aphidicolin; D012313:RNA; D011494:Protein Kinases; D000071877:Checkpoint Kinase 1; D017346:Protein Serine-Threonine Kinases; D042846:CDC2-CDC28 Kinases; C495891:Cdk2 protein, Xenopus; D051357:Cyclin-Dependent Kinase 2; D018844:Cyclin-Dependent Kinases; D019915:DNA Primase; D004256:DNA Polymerase I; D004265:DNA Helicases