A conserved MCM single-stranded DNA binding element is essential for replication initiation
The ring-shaped MCM helicase is essential to all phases of DNA replication. The complex loads at replication origins as an inactive double-hexamer encircling duplex DNA. Helicase activation converts this species to two active single hexamers that encircle single-stranded DNA (ssDNA). The molecular d...
| Main Authors: | , , , , |
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| Other Authors: | |
| Format: | Article |
| Language: | en_US |
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eLife Sciences Publications, Ltd.
2014
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| Online Access: | http://hdl.handle.net/1721.1/89402 https://orcid.org/0000-0002-2876-610X |
| _version_ | 1811097511047200768 |
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| author | Froelich, Clifford A. Kang, Sukhyun Epling, Leslie B. Enemark, Eric J. Bell, Stephen P |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology Froelich, Clifford A. Kang, Sukhyun Epling, Leslie B. Enemark, Eric J. Bell, Stephen P |
| author_sort | Froelich, Clifford A. |
| collection | MIT |
| description | The ring-shaped MCM helicase is essential to all phases of DNA replication. The complex loads at replication origins as an inactive double-hexamer encircling duplex DNA. Helicase activation converts this species to two active single hexamers that encircle single-stranded DNA (ssDNA). The molecular details of MCM DNA interactions during these events are unknown. We determined the crystal structure of the Pyrococcus furiosus MCM N-terminal domain hexamer bound to ssDNA and define a conserved MCM-ssDNA binding motif (MSSB). Intriguingly, ssDNA binds the MCM ring interior perpendicular to the central channel with defined polarity. In eukaryotes, the MSSB is conserved in several Mcm2-7 subunits, and MSSB mutant combinations in S. cerevisiae Mcm2-7 are not viable. Mutant Mcm2-7 complexes assemble and are recruited to replication origins, but are defective in helicase loading and activation. Our findings identify an important MCM-ssDNA interaction and suggest it functions during helicase activation to select the strand for translocation. |
| first_indexed | 2024-09-23T17:00:31Z |
| format | Article |
| id | mit-1721.1/89402 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T17:00:31Z |
| publishDate | 2014 |
| publisher | eLife Sciences Publications, Ltd. |
| record_format | dspace |
| spelling | mit-1721.1/894022022-09-29T23:01:47Z A conserved MCM single-stranded DNA binding element is essential for replication initiation Froelich, Clifford A. Kang, Sukhyun Epling, Leslie B. Enemark, Eric J. Bell, Stephen P Massachusetts Institute of Technology. Department of Biology Kang, Sukhyun Bell, Stephen P. The ring-shaped MCM helicase is essential to all phases of DNA replication. The complex loads at replication origins as an inactive double-hexamer encircling duplex DNA. Helicase activation converts this species to two active single hexamers that encircle single-stranded DNA (ssDNA). The molecular details of MCM DNA interactions during these events are unknown. We determined the crystal structure of the Pyrococcus furiosus MCM N-terminal domain hexamer bound to ssDNA and define a conserved MCM-ssDNA binding motif (MSSB). Intriguingly, ssDNA binds the MCM ring interior perpendicular to the central channel with defined polarity. In eukaryotes, the MSSB is conserved in several Mcm2-7 subunits, and MSSB mutant combinations in S. cerevisiae Mcm2-7 are not viable. Mutant Mcm2-7 complexes assemble and are recruited to replication origins, but are defective in helicase loading and activation. Our findings identify an important MCM-ssDNA interaction and suggest it functions during helicase activation to select the strand for translocation. National Institute of General Medical Sciences (U.S.) (R01GM098771) American Lebanese Syrian Associated Charities National Cancer Institute (U.S.) (Cancer Center Support Grant 5 P30 CA021765-32) Howard Hughes Medical Institute National Institute of General Medical Sciences (U.S.) (R01-GM52339) 2014-09-09T19:57:40Z 2014-09-09T19:57:40Z 2014-04 2013-12 Article http://purl.org/eprint/type/JournalArticle 2050-084X http://hdl.handle.net/1721.1/89402 Froelich, Clifford A, Sukhyun Kang, Leslie B Epling, Stephen P Bell, and Eric J Enemark. “A Conserved MCM Single-Stranded DNA Binding Element Is Essential for Replication Initiation.” eLife 3 (April 1, 2014). pp 1-21. https://orcid.org/0000-0002-2876-610X en_US http://dx.doi.org/10.7554/eLife.01993 eLife Creative Commons Attribution http://creativecommons.org/licenses/by/3.0/ application/pdf eLife Sciences Publications, Ltd. eLife Sciences Publications, Ltd. |
| spellingShingle | Froelich, Clifford A. Kang, Sukhyun Epling, Leslie B. Enemark, Eric J. Bell, Stephen P A conserved MCM single-stranded DNA binding element is essential for replication initiation |
| title | A conserved MCM single-stranded DNA binding element is essential for replication initiation |
| title_full | A conserved MCM single-stranded DNA binding element is essential for replication initiation |
| title_fullStr | A conserved MCM single-stranded DNA binding element is essential for replication initiation |
| title_full_unstemmed | A conserved MCM single-stranded DNA binding element is essential for replication initiation |
| title_short | A conserved MCM single-stranded DNA binding element is essential for replication initiation |
| title_sort | conserved mcm single stranded dna binding element is essential for replication initiation |
| url | http://hdl.handle.net/1721.1/89402 https://orcid.org/0000-0002-2876-610X |
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