CDC6 association with the ORC:origin complex / Cul8-RING ubiquitin ligase complex / maintenance of rDNA / Assembly of the ORC complex at the origin of replication / nuclear origin of replication recognition complex / CDK-mediated phosphorylation and removal of Cdc6 / pre-replicative complex assembly involved in nuclear cell cycle DNA replication / Activation of the pre-replicative complex / Orc1 removal from chromatin / nuclear pre-replicative complex ...CDC6 association with the ORC:origin complex / Cul8-RING ubiquitin ligase complex / maintenance of rDNA / Assembly of the ORC complex at the origin of replication / nuclear origin of replication recognition complex / CDK-mediated phosphorylation and removal of Cdc6 / pre-replicative complex assembly involved in nuclear cell cycle DNA replication / Activation of the pre-replicative complex / Orc1 removal from chromatin / nuclear pre-replicative complex / Activation of ATR in response to replication stress / DNA replication preinitiation complex / mitotic DNA replication checkpoint signaling / silent mating-type cassette heterochromatin formation / regulation of DNA-templated DNA replication initiation / regulation of DNA replication / DNA replication origin binding / subtelomeric heterochromatin formation / DNA replication initiation / nucleosome binding / G1/S transition of mitotic cell cycle / 染色体 / chromosome, telomeric region / 細胞分裂 / GTPase activity / chromatin binding / GTP binding / ATP hydrolysis activity / 核質 / ATP binding / 細胞核 / metal ion binding / 細胞質 類似検索 - 分子機能
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)
R01-GM141313
米国
European Research Council (ERC)
ERC-STG-757909
米国
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)
T32-GM008283
米国
引用
ジャーナル: Nat Commun / 年: 2022 タイトル: A mechanism of origin licensing control through autoinhibition of S. cerevisiae ORC·DNA·Cdc6. 著者: Jan Marten Schmidt / Ran Yang / Ashish Kumar / Olivia Hunker / Jan Seebacher / Franziska Bleichert / 要旨: The coordinated action of multiple replicative helicase loading factors is needed for the licensing of replication origins prior to DNA replication. Binding of the Origin Recognition Complex (ORC) to ...The coordinated action of multiple replicative helicase loading factors is needed for the licensing of replication origins prior to DNA replication. Binding of the Origin Recognition Complex (ORC) to DNA initiates the ATP-dependent recruitment of Cdc6, Cdt1 and Mcm2-7 loading, but the structural details for timely ATPase site regulation and for how loading can be impeded by inhibitory signals, such as cyclin-dependent kinase phosphorylation, are unknown. Using cryo-electron microscopy, we have determined several structures of S. cerevisiae ORC·DNA·Cdc6 intermediates at 2.5-2.7 Å resolution. These structures reveal distinct ring conformations of the initiator·co-loader assembly and inactive ATPase site configurations for ORC and Cdc6. The Orc6 N-terminal domain laterally engages the ORC·Cdc6 ring in a manner that is incompatible with productive Mcm2-7 docking, while deletion of this Orc6 region alleviates the CDK-mediated inhibition of Mcm7 recruitment. Our findings support a model in which Orc6 promotes the assembly of an autoinhibited ORC·DNA·Cdc6 intermediate to block origin licensing in response to CDK phosphorylation and to avert DNA re-replication.