Multiple Functions for Mcm2–7 ATPase Motifs during Replication Initiation
The Mcm2–7 replicative helicase is central to all steps of eukaryotic DNA replication. The hexameric ring of Mcm subunits forms six essential ATPases whose contributions to replication initiation remain unclear. Mcm2–7 complexes containing ATPase-motif mutations showed Mcm2–7 ATP binding and hydroly...
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| Format: | Article |
| Language: | en_US |
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Elsevier
2017
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| Online Access: | http://hdl.handle.net/1721.1/109231 https://orcid.org/0000-0003-1736-6343 https://orcid.org/0000-0002-2876-610X |
| _version_ | 1826189989110611968 |
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| author | Kang, Sukhyun Warner, Megan Dobbins Bell, Stephen P |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology Kang, Sukhyun Warner, Megan Dobbins Bell, Stephen P |
| author_sort | Kang, Sukhyun |
| collection | MIT |
| description | The Mcm2–7 replicative helicase is central to all steps of eukaryotic DNA replication. The hexameric ring of Mcm subunits forms six essential ATPases whose contributions to replication initiation remain unclear. Mcm2–7 complexes containing ATPase-motif mutations showed Mcm2–7 ATP binding and hydrolysis are required for helicase loading. Loading-defective Mcm2–7 mutant complexes were defective in initial Mcm2–7 recruitment or Cdt1 release. Comparison with Cdc6 ATPase mutants showed that Cdc6 ATP hydrolysis is not required for helicase loading but instead drives removal of Mcm2–7 complexes that cannot complete loading. A subset of Mcm2–7 ATPase-site mutants completed helicase loading but could not initiate replication. Individual mutants were defective in distinct events during helicase activation, including maintenance of DNA association, recruitment of the GINS helicase activator, and DNA unwinding. Consistent with its heterohexameric structure, our findings show that the six Mcm2–7 ATPase active sites are specialized for different functions during helicase loading and activation. |
| first_indexed | 2024-09-23T08:33:19Z |
| format | Article |
| id | mit-1721.1/109231 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T08:33:19Z |
| publishDate | 2017 |
| publisher | Elsevier |
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| spelling | mit-1721.1/1092312022-09-30T09:35:07Z Multiple Functions for Mcm2–7 ATPase Motifs during Replication Initiation Kang, Sukhyun Warner, Megan Dobbins Bell, Stephen P Massachusetts Institute of Technology. Department of Biology Kang, Sukhyun Warner, Megan Dobbins Bell, Stephen P The Mcm2–7 replicative helicase is central to all steps of eukaryotic DNA replication. The hexameric ring of Mcm subunits forms six essential ATPases whose contributions to replication initiation remain unclear. Mcm2–7 complexes containing ATPase-motif mutations showed Mcm2–7 ATP binding and hydrolysis are required for helicase loading. Loading-defective Mcm2–7 mutant complexes were defective in initial Mcm2–7 recruitment or Cdt1 release. Comparison with Cdc6 ATPase mutants showed that Cdc6 ATP hydrolysis is not required for helicase loading but instead drives removal of Mcm2–7 complexes that cannot complete loading. A subset of Mcm2–7 ATPase-site mutants completed helicase loading but could not initiate replication. Individual mutants were defective in distinct events during helicase activation, including maintenance of DNA association, recruitment of the GINS helicase activator, and DNA unwinding. Consistent with its heterohexameric structure, our findings show that the six Mcm2–7 ATPase active sites are specialized for different functions during helicase loading and activation. United States. National Institutes of Health (GM052339) United States. National Institutes of Health (GM007287) National Science Foundation (U.S.) (1122374) 2017-05-19T19:24:32Z 2017-05-19T19:24:32Z 2014-07 2014-05 Article http://purl.org/eprint/type/JournalArticle 1097-2765 1097-4164 http://hdl.handle.net/1721.1/109231 Kang, Sukhyun; Warner, Megan D. and Bell, Stephen P. “Multiple Functions for Mcm2–7 ATPase Motifs During Replication Initiation.” Molecular Cell 55, no. 5 (September 2014): 655–665 © 2014 Elsevier Inc https://orcid.org/0000-0003-1736-6343 https://orcid.org/0000-0002-2876-610X en_US http://dx.doi.org/10.1016/j.molcel.2014.06.033 Molecular Cell Creative Commons Attribution-NonCommercial-NoDerivs License http://creativecommons.org/licenses/by-nc-nd/4.0/ application/pdf Elsevier PMC |
| spellingShingle | Kang, Sukhyun Warner, Megan Dobbins Bell, Stephen P Multiple Functions for Mcm2–7 ATPase Motifs during Replication Initiation |
| title | Multiple Functions for Mcm2–7 ATPase Motifs during Replication Initiation |
| title_full | Multiple Functions for Mcm2–7 ATPase Motifs during Replication Initiation |
| title_fullStr | Multiple Functions for Mcm2–7 ATPase Motifs during Replication Initiation |
| title_full_unstemmed | Multiple Functions for Mcm2–7 ATPase Motifs during Replication Initiation |
| title_short | Multiple Functions for Mcm2–7 ATPase Motifs during Replication Initiation |
| title_sort | multiple functions for mcm2 7 atpase motifs during replication initiation |
| url | http://hdl.handle.net/1721.1/109231 https://orcid.org/0000-0003-1736-6343 https://orcid.org/0000-0002-2876-610X |
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