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Mechanistic insights into Lhr helicase function in DNA repair
The DNA helicase Large helicase-related (Lhr) is present throughout archaea, including in the Asgard and Nanoarchaea, and has homologues in bacteria and eukaryotes. It is thought to function in DNA repair but in a context that is not known. Our data show that archaeal Lhr preferentially targets DNA...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Portland Press Ltd.
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7437997/ https://www.ncbi.nlm.nih.gov/pubmed/32706021 http://dx.doi.org/10.1042/BCJ20200379 |
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author | Buckley, Ryan J. Kramm, Kevin Cooper, Christopher D. O. Grohmann, Dina Bolt, Edward L. |
author_facet | Buckley, Ryan J. Kramm, Kevin Cooper, Christopher D. O. Grohmann, Dina Bolt, Edward L. |
author_sort | Buckley, Ryan J. |
collection | PubMed |
description | The DNA helicase Large helicase-related (Lhr) is present throughout archaea, including in the Asgard and Nanoarchaea, and has homologues in bacteria and eukaryotes. It is thought to function in DNA repair but in a context that is not known. Our data show that archaeal Lhr preferentially targets DNA replication fork structures. In a genetic assay, expression of archaeal Lhr gave a phenotype identical to the replication-coupled DNA repair enzymes Hel308 and RecQ. Purified archaeal Lhr preferentially unwound model forked DNA substrates compared with DNA duplexes, flaps and Holliday junctions, and unwound them with directionality. Single-molecule FRET measurements showed that binding of Lhr to a DNA fork causes ATP-independent distortion and base-pair melting at, or close to, the fork branchpoint. ATP-dependent directional translocation of Lhr resulted in fork DNA unwinding through the ‘parental’ DNA strands. Interaction of Lhr with replication forks in vivo and in vitro suggests that it contributes to DNA repair at stalled or broken DNA replication. |
format | Online Article Text |
id | pubmed-7437997 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Portland Press Ltd. |
record_format | MEDLINE/PubMed |
spelling | pubmed-74379972020-08-24 Mechanistic insights into Lhr helicase function in DNA repair Buckley, Ryan J. Kramm, Kevin Cooper, Christopher D. O. Grohmann, Dina Bolt, Edward L. Biochem J DNA, Chromosomes & Chromosomal Structure The DNA helicase Large helicase-related (Lhr) is present throughout archaea, including in the Asgard and Nanoarchaea, and has homologues in bacteria and eukaryotes. It is thought to function in DNA repair but in a context that is not known. Our data show that archaeal Lhr preferentially targets DNA replication fork structures. In a genetic assay, expression of archaeal Lhr gave a phenotype identical to the replication-coupled DNA repair enzymes Hel308 and RecQ. Purified archaeal Lhr preferentially unwound model forked DNA substrates compared with DNA duplexes, flaps and Holliday junctions, and unwound them with directionality. Single-molecule FRET measurements showed that binding of Lhr to a DNA fork causes ATP-independent distortion and base-pair melting at, or close to, the fork branchpoint. ATP-dependent directional translocation of Lhr resulted in fork DNA unwinding through the ‘parental’ DNA strands. Interaction of Lhr with replication forks in vivo and in vitro suggests that it contributes to DNA repair at stalled or broken DNA replication. Portland Press Ltd. 2020-08-28 2020-08-19 /pmc/articles/PMC7437997/ /pubmed/32706021 http://dx.doi.org/10.1042/BCJ20200379 Text en © 2020 The Author(s) https://creativecommons.org/licenses/by/4.0/ This is an open access article published by Portland Press Limited on behalf of the Biochemical Society and distributed under the Creative Commons Attribution License 4.0 (CC BY) (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | DNA, Chromosomes & Chromosomal Structure Buckley, Ryan J. Kramm, Kevin Cooper, Christopher D. O. Grohmann, Dina Bolt, Edward L. Mechanistic insights into Lhr helicase function in DNA repair |
title | Mechanistic insights into Lhr helicase function in DNA repair |
title_full | Mechanistic insights into Lhr helicase function in DNA repair |
title_fullStr | Mechanistic insights into Lhr helicase function in DNA repair |
title_full_unstemmed | Mechanistic insights into Lhr helicase function in DNA repair |
title_short | Mechanistic insights into Lhr helicase function in DNA repair |
title_sort | mechanistic insights into lhr helicase function in dna repair |
topic | DNA, Chromosomes & Chromosomal Structure |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7437997/ https://www.ncbi.nlm.nih.gov/pubmed/32706021 http://dx.doi.org/10.1042/BCJ20200379 |
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