dc.contributor.author | On, KF | |
dc.contributor.author | Beuron, F | |
dc.contributor.author | Frith, D | |
dc.contributor.author | Snijders, AP | |
dc.contributor.author | Morris, EP | |
dc.contributor.author | Diffley, JFX | |
dc.date.accessioned | 2020-08-14T15:39:32Z | |
dc.date.issued | 2014-03-18 | |
dc.identifier.citation | The EMBO journal, 2014, 33 (6), pp. 605 - 620 | |
dc.identifier.issn | 0261-4189 | |
dc.identifier.uri | https://repository.icr.ac.uk/handle/internal/3970 | |
dc.identifier.eissn | 1460-2075 | |
dc.identifier.doi | 10.1002/embj.201387369 | |
dc.description.abstract | Eukaryotic 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. | |
dc.format | Print-Electronic | |
dc.format.extent | 605 - 620 | |
dc.language | eng | |
dc.language.iso | eng | |
dc.publisher | WILEY-BLACKWELL | |
dc.rights.uri | https://creativecommons.org/licenses/by-nc-nd/4.0 | |
dc.subject | Saccharomycetales | |
dc.subject | Multiprotein Complexes | |
dc.subject | Protein-Serine-Threonine Kinases | |
dc.subject | Cell Cycle Proteins | |
dc.subject | Saccharomyces cerevisiae Proteins | |
dc.subject | DNA Replication | |
dc.subject | Enzyme Activation | |
dc.subject | Phosphorylation | |
dc.subject | Replication Origin | |
dc.subject | Plasmids | |
dc.subject | Models, Biological | |
dc.subject | Models, Molecular | |
dc.subject | Mass Spectrometry | |
dc.subject | Minichromosome Maintenance Proteins | |
dc.title | Prereplicative complexes assembled in vitro support origin-dependent and independent DNA replication. | |
dc.type | Journal Article | |
rioxxterms.versionofrecord | 10.1002/embj.201387369 | |
rioxxterms.licenseref.uri | https://creativecommons.org/licenses/by-nc-nd/4.0 | |
rioxxterms.licenseref.startdate | 2014-03 | |
rioxxterms.type | Journal Article/Review | |
dc.relation.isPartOf | The EMBO journal | |
pubs.issue | 6 | |
pubs.notes | Not known | |
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.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-status | Published | |
pubs.volume | 33 | |
pubs.embargo.terms | Not known | |
icr.researchteam | Structural Electron Microscopy | |
dc.contributor.icrauthor | Beuron, Fabienne | |
dc.contributor.icrauthor | Morris, Edward | |