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Bacillus subtilis RarA modulates replication restart
The ubiquitous RarA/Mgs1/WRNIP protein plays a crucial, but poorly understood role in genome maintenance. We show that Bacillus subtilis RarA, in the apo form, preferentially binds single-stranded (ss) over double-stranded (ds) DNA. SsbA bound to ssDNA loads RarA, and for such recruitment the amphip...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6101539/ https://www.ncbi.nlm.nih.gov/pubmed/29947798 http://dx.doi.org/10.1093/nar/gky541 |
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author | Carrasco, Begoña Seco, Elena M López-Sanz, María Alonso, Juan C Ayora, Silvia |
author_facet | Carrasco, Begoña Seco, Elena M López-Sanz, María Alonso, Juan C Ayora, Silvia |
author_sort | Carrasco, Begoña |
collection | PubMed |
description | The ubiquitous RarA/Mgs1/WRNIP protein plays a crucial, but poorly understood role in genome maintenance. We show that Bacillus subtilis RarA, in the apo form, preferentially binds single-stranded (ss) over double-stranded (ds) DNA. SsbA bound to ssDNA loads RarA, and for such recruitment the amphipathic C-terminal domain of SsbA is required. RarA is a DNA-dependent ATPase strongly stimulated by ssDNA–dsDNA junctions and SsbA, or by dsDNA ends. RarA, which may interact with PriA, does not stimulate PriA DNA unwinding. In a reconstituted PriA-dependent DNA replication system, RarA inhibited initiation, but not chain elongation. The RarA effect was not observed in the absence of SsbA, or when the host-encoded preprimosome and the DNA helicase are replaced by proteins from the SPP1 phage with similar function. We propose that RarA assembles at blocked forks to maintain genome integrity. Through its interaction with SsbA and with a preprimosomal component, RarA might impede the assembly of the replicative helicase, to prevent that recombination intermediates contribute to pathological DNA replication restart. |
format | Online Article Text |
id | pubmed-6101539 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-61015392018-08-27 Bacillus subtilis RarA modulates replication restart Carrasco, Begoña Seco, Elena M López-Sanz, María Alonso, Juan C Ayora, Silvia Nucleic Acids Res Genome Integrity, Repair and Replication The ubiquitous RarA/Mgs1/WRNIP protein plays a crucial, but poorly understood role in genome maintenance. We show that Bacillus subtilis RarA, in the apo form, preferentially binds single-stranded (ss) over double-stranded (ds) DNA. SsbA bound to ssDNA loads RarA, and for such recruitment the amphipathic C-terminal domain of SsbA is required. RarA is a DNA-dependent ATPase strongly stimulated by ssDNA–dsDNA junctions and SsbA, or by dsDNA ends. RarA, which may interact with PriA, does not stimulate PriA DNA unwinding. In a reconstituted PriA-dependent DNA replication system, RarA inhibited initiation, but not chain elongation. The RarA effect was not observed in the absence of SsbA, or when the host-encoded preprimosome and the DNA helicase are replaced by proteins from the SPP1 phage with similar function. We propose that RarA assembles at blocked forks to maintain genome integrity. Through its interaction with SsbA and with a preprimosomal component, RarA might impede the assembly of the replicative helicase, to prevent that recombination intermediates contribute to pathological DNA replication restart. Oxford University Press 2018-08-21 2018-06-27 /pmc/articles/PMC6101539/ /pubmed/29947798 http://dx.doi.org/10.1093/nar/gky541 Text en © The Author(s) 2018. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by-nc/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com |
spellingShingle | Genome Integrity, Repair and Replication Carrasco, Begoña Seco, Elena M López-Sanz, María Alonso, Juan C Ayora, Silvia Bacillus subtilis RarA modulates replication restart |
title |
Bacillus subtilis RarA modulates replication restart |
title_full |
Bacillus subtilis RarA modulates replication restart |
title_fullStr |
Bacillus subtilis RarA modulates replication restart |
title_full_unstemmed |
Bacillus subtilis RarA modulates replication restart |
title_short |
Bacillus subtilis RarA modulates replication restart |
title_sort | bacillus subtilis rara modulates replication restart |
topic | Genome Integrity, Repair and Replication |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6101539/ https://www.ncbi.nlm.nih.gov/pubmed/29947798 http://dx.doi.org/10.1093/nar/gky541 |
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