{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"submitter":["Mukherjee PP"],"funding":["Medical Research Council","Wellcome Trust"],"pagination":["2777-2783.e4"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC6899518"],"repository":["biostudies-literature"],"omics_type":["Unknown"],"volume":["28(11)"],"pubmed_abstract":["Disassembly of the replisome is the final step of chromosome duplication in eukaryotes. In budding yeast and metazoa, cullin ubiquitin ligases are required to ubiquitylate the Cdc45-MCM-GINS (CMG) helicase that lies at the heart of the replisome, leading to a disassembly reaction that is dependent upon the ATPase known as Cdc48 or p97. Here, we describe the reconstitution of replisome disassembly, using a purified complex of the budding yeast replisome in association with the cullin ligase SCF<sup>Dia2</sup>. Upon addition of E1 and E2 enzymes, together with ubiquitin and ATP, the CMG helicase is ubiquitylated on its Mcm7 subunit. Subsequent addition of Cdc48, together with its cofactors Ufd1-Npl4, drives efficient disassembly of ubiquitylated CMG, thereby recapitulating the steps of repli"],"journal":["Cell reports"],"pubmed_title":["In Vitro Reconstitution Defines the Minimal Requirements for Cdc48-Dependent Disassembly of the CMG Helicase in Budding Yeast."],"pmcid":["PMC6899518"],"funding_grant_id":["MC_UU_00018/4","MC_UU_12016/13","102943/Z/13/Z"],"pubmed_authors":["Mukherjee PP","Labib KPM"],"additional_accession":[]},"is_claimable":false,"name":"In Vitro Reconstitution Defines the Minimal Requirements for Cdc48-Dependent Disassembly of the CMG Helicase in Budding Yeast.","description":"Disassembly of the replisome is the final step of chromosome duplication in eukaryotes. In budding yeast and metazoa, cullin ubiquitin ligases are required to ubiquitylate the Cdc45-MCM-GINS (CMG) helicase that lies at the heart of the replisome, leading to a disassembly reaction that is dependent upon the ATPase known as Cdc48 or p97. Here, we describe the reconstitution of replisome disassembly, using a purified complex of the budding yeast replisome in association with the cullin ligase SCF<sup>Dia2</sup>. Upon addition of E1 and E2 enzymes, together with ubiquitin and ATP, the CMG helicase is ubiquitylated on its Mcm7 subunit. Subsequent addition of Cdc48, together with its cofactors Ufd1-Npl4, drives efficient disassembly of ubiquitylated CMG, thereby recapitulating the steps of repli","dates":{"release":"2019-01-01T00:00:00Z","publication":"2019 Sep","modification":"2025-04-04T14:59:49.227Z","creation":"2025-04-04T14:59:49.227Z"},"accession":"S-EPMC6899518","cross_references":{"pubmed":["31509741"],"doi":["10.1016/j.celrep.2019.08.026"]}}