title: ATPase-dependent quality control of DNA replication origin licensing.
abstract: The regulated loading of the Mcm2-7 DNA helicase (comprising six related subunits, Mcm2 to Mcm7) into pre-replicative complexes at multiple replication origins ensures precise once per cell cycle replication in eukaryotic cells. The origin recognition complex (ORC), Cdc6 and Cdt1 load Mcm2-7 into a double hexamer bound around duplex DNA in an ATP-dependent reaction, but the molecular mechanism of this origin 'licensing' is still poorly understood. Here we show that both Mcm2-7 hexamers in Saccharomyces cerevisiae are recruited to origins by an essential, conserved carboxy-terminal domain of Mcm3 that interacts with and stimulates the ATPase activity of ORC-Cdc6. ATP hydrolysis can promote Mcm2-7 loading, but can also promote Mcm2-7 release if components are missing or if ORC has been inactivated by cyclin-dependent kinase phosphorylation. Our work provides new insights into how origins are licensed and reveals a novel ATPase-dependent mechanism contributing to precise once per cell cycle replication.
authors.affiliation: Cancer Research UK London Research Institute, Clare Hall Laboratories, South Mimms EN6 3LD, UK.
authors.forename: Jordi
authors.initials: J
authors.lastname: Frigola
authors.forename: Dirk
authors.initials: D
authors.lastname: Remus
authors.forename: Amina
authors.initials: A
authors.lastname: Mehanna
authors.forename: John F X
authors.initials: JF
authors.lastname: Diffley
mesh_terms: D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism; D000595:Amino Acid Sequence; D018797:Cell Cycle Proteins / Q000378:metabolism; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism; D004261:DNA Replication / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000378:metabolism; D004789:Enzyme Activation; D006868:Hydrolysis; D064126:Minichromosome Maintenance Complex Component 3; D064168:Minichromosome Maintenance Complex Component 7; D009687:Nuclear Proteins / Q000378:metabolism; D011485:Protein Binding; D018741:Replication Origin / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism; D016415:Sequence Alignment
chemical_list: D018797:Cell Cycle Proteins; D002868:Chromosomal Proteins, Non-Histone; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D029701:Saccharomyces cerevisiae Proteins; C499592:TAH11 protein, S cerevisiae; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases; C069199:MCM2 protein, S cerevisiae; C507761:MCM3 protein, S cerevisiae; C491088:MCM7 protein, S cerevisiae; D064126:Minichromosome Maintenance Complex Component 3; D064168:Minichromosome Maintenance Complex Component 7