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dc.contributor.authorOn, KFen_US
dc.contributor.authorBeuron, Fen_US
dc.contributor.authorFrith, Den_US
dc.contributor.authorSnijders, APen_US
dc.contributor.authorMorris, EPen_US
dc.contributor.authorDiffley, JFXen_US
dc.date.accessioned2020-08-14T15:39:32Z
dc.date.issued2014-03en_US
dc.identifier.citationThe EMBO journal, 2014, 33 (6), pp. 605 - 620en_US
dc.identifier.issn0261-4189en_US
dc.identifier.urihttps://repository.icr.ac.uk/handle/internal/3970
dc.identifier.eissn1460-2075en_US
dc.identifier.doi10.1002/embj.201387369en_US
dc.description.abstractEukaryotic DNA replication initiates from multiple replication origins. To ensure each origin fires just once per cell cycle, initiation is divided into two biochemically discrete steps: the Mcm2-7 helicase is first loaded into prereplicative complexes (pre-RCs) as an inactive double hexamer by the origin recognition complex (ORC), Cdt1 and Cdc6; the helicase is then activated by a set of "firing factors." Here, we show that plasmids containing pre-RCs assembled with purified proteins support complete and semi-conservative replication in extracts from budding yeast cells overexpressing firing factors. Replication requires cyclin-dependent kinase (CDK) and Dbf4-dependent kinase (DDK). DDK phosphorylation of Mcm2-7 does not by itself promote separation of the double hexamer, but is required for the recruitment of firing factors and replisome components in the extract. Plasmid replication does not require a functional replication origin; however, in the presence of competitor DNA and limiting ORC concentrations, replication becomes origin-dependent in this system. These experiments indicate that Mcm2-7 double hexamers can be precursors of replication and provide insight into the nature of eukaryotic DNA replication origins.en_US
dc.formatPrint-Electronicen_US
dc.format.extent605 - 620en_US
dc.languageengen_US
dc.language.isoengen_US
dc.rights.urihttps://creativecommons.org/licenses/by-nc-nd/4.0/en_US
dc.subjectSaccharomycetalesen_US
dc.subjectMultiprotein Complexesen_US
dc.subjectProtein-Serine-Threonine Kinasesen_US
dc.subjectCell Cycle Proteinsen_US
dc.subjectSaccharomyces cerevisiae Proteinsen_US
dc.subjectDNA Replicationen_US
dc.subjectEnzyme Activationen_US
dc.subjectPhosphorylationen_US
dc.subjectReplication Originen_US
dc.subjectPlasmidsen_US
dc.subjectModels, Biologicalen_US
dc.subjectModels, Molecularen_US
dc.subjectMass Spectrometryen_US
dc.subjectMinichromosome Maintenance Proteinsen_US
dc.titlePrereplicative complexes assembled in vitro support origin-dependent and independent DNA replication.en_US
dc.typeJournal Article
rioxxterms.versionofrecord10.1002/embj.201387369en_US
rioxxterms.licenseref.urihttps://creativecommons.org/licenses/by-nc-nd/4.0en_US
rioxxterms.licenseref.startdate2014-03en_US
rioxxterms.typeJournal Article/Reviewen_US
dc.relation.isPartOfThe EMBO journalen_US
pubs.issue6en_US
pubs.notesNot knownen_US
pubs.organisational-group/ICR
pubs.organisational-group/ICR/Primary Group
pubs.organisational-group/ICR/Primary Group/ICR Divisions
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Structural Biology
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Structural Biology/Structural Electron Microscopy
pubs.publication-statusPublisheden_US
pubs.volume33en_US
pubs.embargo.termsNot knownen_US
icr.researchteamStructural Electron Microscopyen_US
dc.contributor.icrauthorBeuron, Fabienneen_US
dc.contributor.icrauthorMorris, Edwarden_US


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