- EMDB-44441: Cryo-EM structure of the S. cerevisiae ORC-Cdc6-Mcm2-7-DNA comple... -
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基本情報
登録情報
データベース: EMDB / ID: EMD-44441
タイトル
Cryo-EM structure of the S. cerevisiae ORC-Cdc6-Mcm2-7-DNA complex with a fully closed Mcm2-Mcm5 DNA entry gate
マップデータ
structure of the S. cerevisiae ORC-Cdc6-Mcm2-7-DNA complex with a fully closed Mcm2-Mcm5 DNA entry gate
試料
複合体: ORC-Cdc6-Mcm2-7-dsDNA
タンパク質・ペプチド: x 14種
DNA: x 2種
リガンド: x 2種
キーワード
DNA replication / Cryo-EM / OCCM-deltaC6 / REPLICATION-DNA complex / DNA BINDING PROTEIN
機能・相同性
機能・相同性情報
MCM complex loading / : / CDC6 association with the ORC:origin complex / replication fork protection complex / Cul8-RING ubiquitin ligase complex / maintenance of rDNA / MCM core complex / Assembly of the pre-replicative complex / Switching of origins to a post-replicative state / MCM complex binding ...MCM complex loading / : / CDC6 association with the ORC:origin complex / replication fork protection complex / Cul8-RING ubiquitin ligase complex / maintenance of rDNA / MCM core complex / Assembly of the pre-replicative complex / Switching of origins to a post-replicative state / MCM complex binding / nuclear DNA replication / premeiotic DNA replication / Assembly of the ORC complex at the origin of replication / pre-replicative complex assembly involved in nuclear cell cycle DNA replication / nuclear origin of replication recognition complex / Activation of the pre-replicative complex / mitotic DNA replication / CMG complex / nuclear pre-replicative complex / cyclin-dependent protein serine/threonine kinase inhibitor activity / nucleosome organization / DNA replication preinitiation complex / Activation of ATR in response to replication stress / mitotic DNA replication checkpoint signaling / double-strand break repair via break-induced replication / MCM complex / mitotic DNA replication initiation / silent mating-type cassette heterochromatin formation / single-stranded DNA helicase activity / regulation of DNA-templated DNA replication initiation / DNA strand elongation involved in DNA replication / CDK-mediated phosphorylation and removal of Cdc6 / Orc1 removal from chromatin / nuclear replication fork / 3'-5' DNA helicase activity / regulation of DNA replication / DNA replication origin binding / DNA replication initiation / subtelomeric heterochromatin formation / nucleosome binding / DNA helicase activity / G1/S transition of mitotic cell cycle / peroxisome / single-stranded DNA binding / chromosome / DNA helicase / DNA replication / chromosome, telomeric region / cell division / hydrolase activity / GTPase activity / DNA damage response / chromatin binding / GTP binding / ATP hydrolysis activity / zinc ion binding / nucleoplasm / ATP binding / metal ion binding / nucleus / cytoplasm 類似検索 - 分子機能
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)
GM110387
米国
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)
GM131754
米国
Biotechnology and Biological Sciences Research Council (BBSRC)
BB/S001387/1
英国
Biotechnology and Biological Sciences Research Council (BBSRC)
BB/N000323/1
英国
Wellcome Trust
107903/Z/15/Z
英国
引用
ジャーナル: Nat Commun / 年: 2025 タイトル: MCM2-7 ring closure involves the Mcm5 C-terminus and triggers Mcm4 ATP hydrolysis. 著者: Sarah V Faull / Marta Barbon / Audrey Mossler / Zuanning Yuan / Lin Bai / L Maximilian Reuter / Alberto Riera / Christian Winkler / Indiana Magdalou / Matthew Peach / Huilin Li / Christian Speck / 要旨: The eukaryotic helicase MCM2-7, is loaded by ORC, Cdc6 and Cdt1 as a double-hexamer onto replication origins. The insertion of DNA into the helicase leads to partial MCM2-7 ring closure, while ATP ...The eukaryotic helicase MCM2-7, is loaded by ORC, Cdc6 and Cdt1 as a double-hexamer onto replication origins. The insertion of DNA into the helicase leads to partial MCM2-7 ring closure, while ATP hydrolysis is essential for consecutive steps in pre-replicative complex (pre-RC) assembly. Currently it is unknown how MCM2-7 ring closure and ATP-hydrolysis are controlled. A cryo-EM structure of an ORC-Cdc6-Cdt1-MCM2-7 intermediate shows a remodelled, fully-closed Mcm2/Mcm5 interface. The Mcm5 C-terminus (C5) contacts Orc3 and specifically recognises this closed ring. Interestingly, we found that normal helicase loading triggers Mcm4 ATP-hydrolysis, which in turn leads to reorganisation of the MCM2-7 complex and Cdt1 release. However, defective MCM2-7 ring closure, due to mutations at the Mcm2/Mcm5 interface, leads to MCM2-7 ring splitting and complex disassembly. As such we identify Mcm4 as the key ATPase in regulating pre-RC formation. Crucially, a stable Mcm2/Mcm5 interface is essential for productive ATP-hydrolysis-dependent remodelling of the helicase.