DDK regulates replication initiation by controlling the multiplicity of Cdc45-GINS binding to Mcm2-7
© 2021, eLife Sciences Publications Ltd. All rights reserved. The committed step of eukaryotic DNA replication occurs when the pairs of Mcm2-7 replicative helicases that license each replication origin are activated. Helicase activation requires the recruitment of Cdc45 and GINS to Mcm2-7, forming C...
| Main Authors: | , , , , , |
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| Other Authors: | |
| Format: | Article |
| Language: | English |
| Published: |
eLife Sciences Publications, Ltd
2021
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| Online Access: | https://hdl.handle.net/1721.1/133327 |
| _version_ | 1811076521715040256 |
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| author | De Jesús-Kim, Lorraine Friedman, Larry J Lõoke, Marko Ramsoomair, Christian K Gelles, Jeff Bell, Stephen P |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology De Jesús-Kim, Lorraine Friedman, Larry J Lõoke, Marko Ramsoomair, Christian K Gelles, Jeff Bell, Stephen P |
| author_sort | De Jesús-Kim, Lorraine |
| collection | MIT |
| description | © 2021, eLife Sciences Publications Ltd. All rights reserved. The committed step of eukaryotic DNA replication occurs when the pairs of Mcm2-7 replicative helicases that license each replication origin are activated. Helicase activation requires the recruitment of Cdc45 and GINS to Mcm2-7, forming Cdc45-Mcm2-7-GINS complexes (CMGs). Using single-molecule biochemical assays to monitor CMG formation, we found that Cdc45 and GINS are recruited to loaded Mcm2-7 in two stages. Initially, Cdc45, GINS, and likely additional proteins are recruited to unstructured Mcm2-7 N-terminal tails in a Dbf4-dependent kinase (DDK)-dependent manner, forming Cdc45-tail-GINS intermediates (CtGs). DDK phosphorylation of multiple phosphorylation sites on the Mcm2-7 tails modulates the number of CtGs formed per Mcm2-7. In a second, inefficient event, a subset of CtGs transfer their Cdc45 and GINS components to form CMGs. Importantly, higher CtG multiplicity increases the frequency of CMG formation. Our findings reveal molecular mechanisms sensitizing helicase activation to DDK levels with implications for control of replication origin efficiency and timing. |
| first_indexed | 2024-09-23T10:23:30Z |
| format | Article |
| id | mit-1721.1/133327 |
| institution | Massachusetts Institute of Technology |
| language | English |
| last_indexed | 2024-09-23T10:23:30Z |
| publishDate | 2021 |
| publisher | eLife Sciences Publications, Ltd |
| record_format | dspace |
| spelling | mit-1721.1/1333272023-12-13T15:09:28Z DDK regulates replication initiation by controlling the multiplicity of Cdc45-GINS binding to Mcm2-7 De Jesús-Kim, Lorraine Friedman, Larry J Lõoke, Marko Ramsoomair, Christian K Gelles, Jeff Bell, Stephen P Massachusetts Institute of Technology. Department of Biology Howard Hughes Medical Institute © 2021, eLife Sciences Publications Ltd. All rights reserved. The committed step of eukaryotic DNA replication occurs when the pairs of Mcm2-7 replicative helicases that license each replication origin are activated. Helicase activation requires the recruitment of Cdc45 and GINS to Mcm2-7, forming Cdc45-Mcm2-7-GINS complexes (CMGs). Using single-molecule biochemical assays to monitor CMG formation, we found that Cdc45 and GINS are recruited to loaded Mcm2-7 in two stages. Initially, Cdc45, GINS, and likely additional proteins are recruited to unstructured Mcm2-7 N-terminal tails in a Dbf4-dependent kinase (DDK)-dependent manner, forming Cdc45-tail-GINS intermediates (CtGs). DDK phosphorylation of multiple phosphorylation sites on the Mcm2-7 tails modulates the number of CtGs formed per Mcm2-7. In a second, inefficient event, a subset of CtGs transfer their Cdc45 and GINS components to form CMGs. Importantly, higher CtG multiplicity increases the frequency of CMG formation. Our findings reveal molecular mechanisms sensitizing helicase activation to DDK levels with implications for control of replication origin efficiency and timing. 2021-10-27T19:52:09Z 2021-10-27T19:52:09Z 2021 2021-07-14T15:05:42Z Article http://purl.org/eprint/type/JournalArticle https://hdl.handle.net/1721.1/133327 en 10.7554/eLife.65471 eLife Creative Commons Attribution 4.0 International license https://creativecommons.org/licenses/by/4.0/ application/pdf eLife Sciences Publications, Ltd eLife |
| spellingShingle | De Jesús-Kim, Lorraine Friedman, Larry J Lõoke, Marko Ramsoomair, Christian K Gelles, Jeff Bell, Stephen P DDK regulates replication initiation by controlling the multiplicity of Cdc45-GINS binding to Mcm2-7 |
| title | DDK regulates replication initiation by controlling the multiplicity of Cdc45-GINS binding to Mcm2-7 |
| title_full | DDK regulates replication initiation by controlling the multiplicity of Cdc45-GINS binding to Mcm2-7 |
| title_fullStr | DDK regulates replication initiation by controlling the multiplicity of Cdc45-GINS binding to Mcm2-7 |
| title_full_unstemmed | DDK regulates replication initiation by controlling the multiplicity of Cdc45-GINS binding to Mcm2-7 |
| title_short | DDK regulates replication initiation by controlling the multiplicity of Cdc45-GINS binding to Mcm2-7 |
| title_sort | ddk regulates replication initiation by controlling the multiplicity of cdc45 gins binding to mcm2 7 |
| url | https://hdl.handle.net/1721.1/133327 |
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