Mcm10 regulates DNA replication elongation by stimulating the CMG replicative helicase
Activation of the Mcm2–7 replicative DNA helicase is the committed step in eukaryotic DNA replication initiation. Although Mcm2–7 activation requires binding of the helicase-activating proteins Cdc45 and GINS (forming the CMG complex), an additional protein, Mcm10, drives initial origin DNA unwindin...
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| Format: | Article |
| Language: | en_US |
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Cold Spring Harbor Laboratory Press
2017
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| Online Access: | http://hdl.handle.net/1721.1/111151 https://orcid.org/0000-0002-1017-2936 https://orcid.org/0000-0002-0559-961X https://orcid.org/0000-0002-2876-610X |
| _version_ | 1826216695322116096 |
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| author | Looke, Marko Maloney, Michael Finnan Bell, Stephen P |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology Looke, Marko Maloney, Michael Finnan Bell, Stephen P |
| author_sort | Looke, Marko |
| collection | MIT |
| description | Activation of the Mcm2–7 replicative DNA helicase is the committed step in eukaryotic DNA replication initiation. Although Mcm2–7 activation requires binding of the helicase-activating proteins Cdc45 and GINS (forming the CMG complex), an additional protein, Mcm10, drives initial origin DNA unwinding by an unknown mechanism. We show that Mcm10 binds a conserved motif located between the oligonucleotide/oligosaccharide fold (OB-fold) and A subdomain of Mcm2. Although buried in the interface between these domains in Mcm2–7 structures, mutations predicted to separate the domains and expose this motif restore growth to conditional-lethal MCM10 mutant cells. We found that, in addition to stimulating initial DNA unwinding, Mcm10 stabilizes Cdc45 and GINS association with Mcm2–7 and stimulates replication elongation in vivo and in vitro. Furthermore, we identified a lethal allele of MCM10 that stimulates initial DNA unwinding but is defective in replication elongation and CMG binding. Our findings expand the roles of Mcm10 during DNA replication and suggest a new model for Mcm10 function as an activator of the CMG complex throughout DNA replication. |
| first_indexed | 2024-09-23T16:51:56Z |
| format | Article |
| id | mit-1721.1/111151 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T16:51:56Z |
| publishDate | 2017 |
| publisher | Cold Spring Harbor Laboratory Press |
| record_format | dspace |
| spelling | mit-1721.1/1111512022-10-03T08:48:12Z Mcm10 regulates DNA replication elongation by stimulating the CMG replicative helicase Looke, Marko Maloney, Michael Finnan Bell, Stephen P Massachusetts Institute of Technology. Department of Biology Looke, Marko Maloney, Michael Finnan Bell, Stephen P Activation of the Mcm2–7 replicative DNA helicase is the committed step in eukaryotic DNA replication initiation. Although Mcm2–7 activation requires binding of the helicase-activating proteins Cdc45 and GINS (forming the CMG complex), an additional protein, Mcm10, drives initial origin DNA unwinding by an unknown mechanism. We show that Mcm10 binds a conserved motif located between the oligonucleotide/oligosaccharide fold (OB-fold) and A subdomain of Mcm2. Although buried in the interface between these domains in Mcm2–7 structures, mutations predicted to separate the domains and expose this motif restore growth to conditional-lethal MCM10 mutant cells. We found that, in addition to stimulating initial DNA unwinding, Mcm10 stabilizes Cdc45 and GINS association with Mcm2–7 and stimulates replication elongation in vivo and in vitro. Furthermore, we identified a lethal allele of MCM10 that stimulates initial DNA unwinding but is defective in replication elongation and CMG binding. Our findings expand the roles of Mcm10 during DNA replication and suggest a new model for Mcm10 function as an activator of the CMG complex throughout DNA replication. National Cancer Institute (U.S.) (Grant P30-CA1405) 2017-09-07T16:31:34Z 2017-09-07T16:31:34Z 2017-02 2016-09 Article http://purl.org/eprint/type/JournalArticle 0890-9369 1549-5477 http://hdl.handle.net/1721.1/111151 Lõoke, Marko, et al. “Mcm10 Regulates DNA Replication Elongation by Stimulating the CMG Replicative Helicase.” Genes & Development 31, 3 (February 2017): 291–305 © 2017 Lõoke et al https://orcid.org/0000-0002-1017-2936 https://orcid.org/0000-0002-0559-961X https://orcid.org/0000-0002-2876-610X en_US http://dx.doi.org/10.1101/gad.291336.116 Genes & Development Creative Commons Attribution-NonCommercial 4.0 International http://creativecommons.org/licenses/by-nc/4.0/ application/pdf Cold Spring Harbor Laboratory Press Cold Spring Harbor Laboratory Press |
| spellingShingle | Looke, Marko Maloney, Michael Finnan Bell, Stephen P Mcm10 regulates DNA replication elongation by stimulating the CMG replicative helicase |
| title | Mcm10 regulates DNA replication elongation by stimulating the CMG replicative helicase |
| title_full | Mcm10 regulates DNA replication elongation by stimulating the CMG replicative helicase |
| title_fullStr | Mcm10 regulates DNA replication elongation by stimulating the CMG replicative helicase |
| title_full_unstemmed | Mcm10 regulates DNA replication elongation by stimulating the CMG replicative helicase |
| title_short | Mcm10 regulates DNA replication elongation by stimulating the CMG replicative helicase |
| title_sort | mcm10 regulates dna replication elongation by stimulating the cmg replicative helicase |
| url | http://hdl.handle.net/1721.1/111151 https://orcid.org/0000-0002-1017-2936 https://orcid.org/0000-0002-0559-961X https://orcid.org/0000-0002-2876-610X |
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