Dna2 nuclease-helicase structure, mechanism and regulation by Rpa
The Dna2 nuclease-helicase maintains genomic integrity by processing DNA double-strand breaks, Okazaki fragments and stalled replication forks. Dna2 requires ssDNA ends, and is dependent on the ssDNA-binding protein Rpa, which controls cleavage polarity. Here we present the 2.3 Å structure of intact...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2015-10-01
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| Series: | eLife |
| Subjects: | |
| Online Access: | https://elifesciences.org/articles/09832 |
| _version_ | 1818019657657876480 |
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| author | Chun Zhou Sergei Pourmal Nikola P Pavletich |
| author_facet | Chun Zhou Sergei Pourmal Nikola P Pavletich |
| author_sort | Chun Zhou |
| collection | DOAJ |
| description | The Dna2 nuclease-helicase maintains genomic integrity by processing DNA double-strand breaks, Okazaki fragments and stalled replication forks. Dna2 requires ssDNA ends, and is dependent on the ssDNA-binding protein Rpa, which controls cleavage polarity. Here we present the 2.3 Å structure of intact mouse Dna2 bound to a 15-nucleotide ssDNA. The nuclease active site is embedded in a long, narrow tunnel through which the DNA has to thread. The helicase domain is required for DNA binding but not threading. We also present the structure of a flexibly-tethered Dna2-Rpa interaction that recruits Dna2 to Rpa-coated DNA. We establish that a second Dna2-Rpa interaction is mutually exclusive with Rpa-DNA interactions and mediates the displacement of Rpa from ssDNA. This interaction occurs at the nuclease tunnel entrance and the 5’ end of the Rpa-DNA complex. Hence, it only displaces Rpa from the 5’ but not 3’ end, explaining how Rpa regulates cleavage polarity. |
| first_indexed | 2024-04-14T07:55:12Z |
| format | Article |
| id | doaj.art-e05ce85e355f43ecbece3e1066f448bf |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-14T07:55:12Z |
| publishDate | 2015-10-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-e05ce85e355f43ecbece3e1066f448bf2022-12-22T02:05:05ZengeLife Sciences Publications LtdeLife2050-084X2015-10-01410.7554/eLife.09832Dna2 nuclease-helicase structure, mechanism and regulation by RpaChun Zhou0Sergei Pourmal1Nikola P Pavletich2Structural Biology Program, Memorial Sloan Kettering Cancer Center, New York, United StatesStructural Biology Program, Memorial Sloan Kettering Cancer Center, New York, United StatesHoward Hughes Medical Institute, Memorial Sloan-Kettering Cancer Center, New York, United StatesThe Dna2 nuclease-helicase maintains genomic integrity by processing DNA double-strand breaks, Okazaki fragments and stalled replication forks. Dna2 requires ssDNA ends, and is dependent on the ssDNA-binding protein Rpa, which controls cleavage polarity. Here we present the 2.3 Å structure of intact mouse Dna2 bound to a 15-nucleotide ssDNA. The nuclease active site is embedded in a long, narrow tunnel through which the DNA has to thread. The helicase domain is required for DNA binding but not threading. We also present the structure of a flexibly-tethered Dna2-Rpa interaction that recruits Dna2 to Rpa-coated DNA. We establish that a second Dna2-Rpa interaction is mutually exclusive with Rpa-DNA interactions and mediates the displacement of Rpa from ssDNA. This interaction occurs at the nuclease tunnel entrance and the 5’ end of the Rpa-DNA complex. Hence, it only displaces Rpa from the 5’ but not 3’ end, explaining how Rpa regulates cleavage polarity.https://elifesciences.org/articles/09832Dna2RpaDNA replicationnucleasehelicaseDNA end resection |
| spellingShingle | Chun Zhou Sergei Pourmal Nikola P Pavletich Dna2 nuclease-helicase structure, mechanism and regulation by Rpa eLife Dna2 Rpa DNA replication nuclease helicase DNA end resection |
| title | Dna2 nuclease-helicase structure, mechanism and regulation by Rpa |
| title_full | Dna2 nuclease-helicase structure, mechanism and regulation by Rpa |
| title_fullStr | Dna2 nuclease-helicase structure, mechanism and regulation by Rpa |
| title_full_unstemmed | Dna2 nuclease-helicase structure, mechanism and regulation by Rpa |
| title_short | Dna2 nuclease-helicase structure, mechanism and regulation by Rpa |
| title_sort | dna2 nuclease helicase structure mechanism and regulation by rpa |
| topic | Dna2 Rpa DNA replication nuclease helicase DNA end resection |
| url | https://elifesciences.org/articles/09832 |
| work_keys_str_mv | AT chunzhou dna2nucleasehelicasestructuremechanismandregulationbyrpa AT sergeipourmal dna2nucleasehelicasestructuremechanismandregulationbyrpa AT nikolappavletich dna2nucleasehelicasestructuremechanismandregulationbyrpa |