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RecG and UvsW catalyse robust DNA rewinding critical for stalled DNA replication fork rescue

Helicases that both unwind and rewind DNA have central roles in DNA repair and genetic recombination. In contrast to unwinding, DNA rewinding by helicases has proved difficult to characterize biochemically because of its thermodynamically downhill nature. Here we use single-molecule assays to mechan...

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Detalles Bibliográficos
Autores principales: Manosas, Maria, Perumal, Senthil K., Bianco, Piero, Ritort, Felix, Benkovic, Stephen J., Croquette, Vincent
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Pub. Group 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3778716/
https://www.ncbi.nlm.nih.gov/pubmed/24013402
http://dx.doi.org/10.1038/ncomms3368
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author Manosas, Maria
Perumal, Senthil K.
Bianco, Piero
Ritort, Felix
Benkovic, Stephen J.
Croquette, Vincent
author_facet Manosas, Maria
Perumal, Senthil K.
Bianco, Piero
Ritort, Felix
Benkovic, Stephen J.
Croquette, Vincent
author_sort Manosas, Maria
collection PubMed
description Helicases that both unwind and rewind DNA have central roles in DNA repair and genetic recombination. In contrast to unwinding, DNA rewinding by helicases has proved difficult to characterize biochemically because of its thermodynamically downhill nature. Here we use single-molecule assays to mechanically destabilize a DNA molecule and follow, in real time, unwinding and rewinding by two DNA repair helicases, bacteriophage T4 UvsW and Escherichia coli RecG. We find that both enzymes are robust rewinding enzymes, which can work against opposing forces as large as 35 pN, revealing their active character. The generation of work during the rewinding reaction allows them to couple rewinding to DNA unwinding and/or protein displacement reactions central to the rescue of stalled DNA replication forks. The overall results support a general mechanism for monomeric rewinding enzymes.
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spelling pubmed-37787162013-09-23 RecG and UvsW catalyse robust DNA rewinding critical for stalled DNA replication fork rescue Manosas, Maria Perumal, Senthil K. Bianco, Piero Ritort, Felix Benkovic, Stephen J. Croquette, Vincent Nat Commun Article Helicases that both unwind and rewind DNA have central roles in DNA repair and genetic recombination. In contrast to unwinding, DNA rewinding by helicases has proved difficult to characterize biochemically because of its thermodynamically downhill nature. Here we use single-molecule assays to mechanically destabilize a DNA molecule and follow, in real time, unwinding and rewinding by two DNA repair helicases, bacteriophage T4 UvsW and Escherichia coli RecG. We find that both enzymes are robust rewinding enzymes, which can work against opposing forces as large as 35 pN, revealing their active character. The generation of work during the rewinding reaction allows them to couple rewinding to DNA unwinding and/or protein displacement reactions central to the rescue of stalled DNA replication forks. The overall results support a general mechanism for monomeric rewinding enzymes. Nature Pub. Group 2013-09-09 /pmc/articles/PMC3778716/ /pubmed/24013402 http://dx.doi.org/10.1038/ncomms3368 Text en Copyright © 2013, Nature Publishing Group, a division of Macmillan Publishers Limited. All Rights Reserved. http://creativecommons.org/licenses/by-nc-by/3.0/ This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. To view a copy of this licence visit http://creativecommons.org/licenses/by/3.0/.
spellingShingle Article
Manosas, Maria
Perumal, Senthil K.
Bianco, Piero
Ritort, Felix
Benkovic, Stephen J.
Croquette, Vincent
RecG and UvsW catalyse robust DNA rewinding critical for stalled DNA replication fork rescue
title RecG and UvsW catalyse robust DNA rewinding critical for stalled DNA replication fork rescue
title_full RecG and UvsW catalyse robust DNA rewinding critical for stalled DNA replication fork rescue
title_fullStr RecG and UvsW catalyse robust DNA rewinding critical for stalled DNA replication fork rescue
title_full_unstemmed RecG and UvsW catalyse robust DNA rewinding critical for stalled DNA replication fork rescue
title_short RecG and UvsW catalyse robust DNA rewinding critical for stalled DNA replication fork rescue
title_sort recg and uvsw catalyse robust dna rewinding critical for stalled dna replication fork rescue
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3778716/
https://www.ncbi.nlm.nih.gov/pubmed/24013402
http://dx.doi.org/10.1038/ncomms3368
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