Mechanism and timing of Mcm2–7 ring closure during DNA replication origin licensing
The opening and closing of two ring-shaped Mcm2-7 DNA helicases is necessary to license eukaryotic origins of replication, although the mechanisms controlling these events are unclear. The origin-recognition complex (ORC), Cdc6 and Cdt1 facilitate this process by establishing a topological link betw...
| Main Authors: | , , , , , |
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| Format: | Article |
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Springer Nature
2018
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| Online Access: | http://hdl.handle.net/1721.1/116464 https://orcid.org/0000-0001-9500-7544 https://orcid.org/0000-0001-8265-4654 https://orcid.org/0000-0002-2876-610X |
| _version_ | 1826200749580746752 |
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| author | Ticau, Simina Friedman, Larry J Champasa, Kanokwan Corrêa, Ivan R Gelles, Jeff Bell, Stephen P |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology Ticau, Simina Friedman, Larry J Champasa, Kanokwan Corrêa, Ivan R Gelles, Jeff Bell, Stephen P |
| author_sort | Ticau, Simina |
| collection | MIT |
| description | The opening and closing of two ring-shaped Mcm2-7 DNA helicases is necessary to license eukaryotic origins of replication, although the mechanisms controlling these events are unclear. The origin-recognition complex (ORC), Cdc6 and Cdt1 facilitate this process by establishing a topological link between each Mcm2-7 hexamer and origin DNA. Using colocalization single-molecule spectroscopy and single-molecule Förster resonance energy transfer (FRET), we monitored ring opening and closing of Saccharomyces cerevisiae Mcm2-7 during origin licensing. The two Mcm2-7 rings were open during initial DNA association and closed sequentially, concomitant with the release of their associated Cdt1. We observed that ATP hydrolysis by Mcm2-7 was coupled to ring closure and Cdt1 release, and failure to load the first Mcm2-7 prevented recruitment of the second Mcm2-7. Our findings identify key mechanisms controlling the Mcm2-7 DNA-entry gate during origin licensing, and reveal that the two Mcm2-7 complexes are loaded via a coordinated series of events with implications for bidirectional replication initiation and quality control. |
| first_indexed | 2024-09-23T11:41:10Z |
| format | Article |
| id | mit-1721.1/116464 |
| institution | Massachusetts Institute of Technology |
| last_indexed | 2024-09-23T11:41:10Z |
| publishDate | 2018 |
| publisher | Springer Nature |
| record_format | dspace |
| spelling | mit-1721.1/1164642022-09-27T21:13:52Z Mechanism and timing of Mcm2–7 ring closure during DNA replication origin licensing Ticau, Simina Friedman, Larry J Champasa, Kanokwan Corrêa, Ivan R Gelles, Jeff Bell, Stephen P Massachusetts Institute of Technology. Department of Biology Ticau, Simina Champasa, Kanokwan Bell, Stephen P The opening and closing of two ring-shaped Mcm2-7 DNA helicases is necessary to license eukaryotic origins of replication, although the mechanisms controlling these events are unclear. The origin-recognition complex (ORC), Cdc6 and Cdt1 facilitate this process by establishing a topological link between each Mcm2-7 hexamer and origin DNA. Using colocalization single-molecule spectroscopy and single-molecule Förster resonance energy transfer (FRET), we monitored ring opening and closing of Saccharomyces cerevisiae Mcm2-7 during origin licensing. The two Mcm2-7 rings were open during initial DNA association and closed sequentially, concomitant with the release of their associated Cdt1. We observed that ATP hydrolysis by Mcm2-7 was coupled to ring closure and Cdt1 release, and failure to load the first Mcm2-7 prevented recruitment of the second Mcm2-7. Our findings identify key mechanisms controlling the Mcm2-7 DNA-entry gate during origin licensing, and reveal that the two Mcm2-7 complexes are loaded via a coordinated series of events with implications for bidirectional replication initiation and quality control. National Institutes of Health (U.S.) (Grant R01 GM52339) National Institutes of Health (U.S.) (Pre-Doctoral Training Grant GM007287) National Cancer Institute (U.S.) (Koch Institute Support Grant P30-CA14051) 2018-06-21T13:50:39Z 2018-06-21T13:50:39Z 2017-03 2018-06-13T17:39:23Z Article http://purl.org/eprint/type/JournalArticle 1545-9993 1545-9985 http://hdl.handle.net/1721.1/116464 Ticau, Simina, et al. “Mechanism and Timing of Mcm2–7 Ring Closure during DNA Replication Origin Licensing.” Nature Structural & Molecular Biology, vol. 24, no. 3, Mar. 2017, pp. 309–15. https://orcid.org/0000-0001-9500-7544 https://orcid.org/0000-0001-8265-4654 https://orcid.org/0000-0002-2876-610X http://dx.doi.org/10.1038/NSMB.3375 Nature Structural & Molecular Biology Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use. application/pdf Springer Nature PMC |
| spellingShingle | Ticau, Simina Friedman, Larry J Champasa, Kanokwan Corrêa, Ivan R Gelles, Jeff Bell, Stephen P Mechanism and timing of Mcm2–7 ring closure during DNA replication origin licensing |
| title | Mechanism and timing of Mcm2–7 ring closure during DNA replication origin licensing |
| title_full | Mechanism and timing of Mcm2–7 ring closure during DNA replication origin licensing |
| title_fullStr | Mechanism and timing of Mcm2–7 ring closure during DNA replication origin licensing |
| title_full_unstemmed | Mechanism and timing of Mcm2–7 ring closure during DNA replication origin licensing |
| title_short | Mechanism and timing of Mcm2–7 ring closure during DNA replication origin licensing |
| title_sort | mechanism and timing of mcm2 7 ring closure during dna replication origin licensing |
| url | http://hdl.handle.net/1721.1/116464 https://orcid.org/0000-0001-9500-7544 https://orcid.org/0000-0001-8265-4654 https://orcid.org/0000-0002-2876-610X |
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