MCM2-7 ring closure involves the Mcm5 C-terminus and triggers Mcm4 ATP hydrolysis
Abstract 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. Cu...
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Nature Portfolio
2025-01-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-024-55479-1 |
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| author | 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 |
| author_facet | 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 |
| author_sort | Sarah V. Faull |
| collection | DOAJ |
| description | Abstract 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. |
| format | Article |
| id | doaj-art-8f45df5e796d4860b6f65dba8fcaaa88 |
| institution | Kabale University |
| issn | 2041-1723 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Nature Portfolio |
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| spelling | doaj-art-8f45df5e796d4860b6f65dba8fcaaa882025-01-05T12:36:58ZengNature PortfolioNature Communications2041-17232025-01-0116111610.1038/s41467-024-55479-1MCM2-7 ring closure involves the Mcm5 C-terminus and triggers Mcm4 ATP hydrolysisSarah V. Faull0Marta Barbon1Audrey Mossler2Zuanning Yuan3Lin Bai4L. Maximilian Reuter5Alberto Riera6Christian Winkler7Indiana Magdalou8Matthew Peach9Huilin Li10Christian Speck11DNA Replication Group, Institute of Clinical Science, Imperial College LondonDNA Replication Group, Institute of Clinical Science, Imperial College LondonDNA Replication Group, Institute of Clinical Science, Imperial College LondonStructural Biology Program, Van Andel Research InstituteStructural Biology Program, Van Andel Research InstituteDNA Replication Group, Institute of Clinical Science, Imperial College LondonDNA Replication Group, Institute of Clinical Science, Imperial College LondonDNA Replication Group, Institute of Clinical Science, Imperial College LondonDNA Replication Group, Institute of Clinical Science, Imperial College LondonDNA Replication Group, Institute of Clinical Science, Imperial College LondonStructural Biology Program, Van Andel Research InstituteDNA Replication Group, Institute of Clinical Science, Imperial College LondonAbstract 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.https://doi.org/10.1038/s41467-024-55479-1 |
| spellingShingle | 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 MCM2-7 ring closure involves the Mcm5 C-terminus and triggers Mcm4 ATP hydrolysis Nature Communications |
| title | MCM2-7 ring closure involves the Mcm5 C-terminus and triggers Mcm4 ATP hydrolysis |
| title_full | MCM2-7 ring closure involves the Mcm5 C-terminus and triggers Mcm4 ATP hydrolysis |
| title_fullStr | MCM2-7 ring closure involves the Mcm5 C-terminus and triggers Mcm4 ATP hydrolysis |
| title_full_unstemmed | MCM2-7 ring closure involves the Mcm5 C-terminus and triggers Mcm4 ATP hydrolysis |
| title_short | MCM2-7 ring closure involves the Mcm5 C-terminus and triggers Mcm4 ATP hydrolysis |
| title_sort | mcm2 7 ring closure involves the mcm5 c terminus and triggers mcm4 atp hydrolysis |
| url | https://doi.org/10.1038/s41467-024-55479-1 |
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