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chi sequences switch the RecBCD helicase–nuclease complex from degradative to replicative modes during the completion of DNA replication

Accurately completing DNA replication when two forks converge is essential to genomic stability. The RecBCD helicase–nuclease complex plays a central role in completion by promoting resection and joining of the excess DNA created when replisomes converge. chi sequences alter RecBCD activity and loca...

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Autores principales: Hamilton, Nicklas A., Jehru, Avery E., Samples, William N., Wendel, Brian M., Mokhtari, Parisa D., Courcelle, Charmain T., Courcelle, Justin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Biochemistry and Molecular Biology 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10025158/
https://www.ncbi.nlm.nih.gov/pubmed/36781123
http://dx.doi.org/10.1016/j.jbc.2023.103013
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author Hamilton, Nicklas A.
Jehru, Avery E.
Samples, William N.
Wendel, Brian M.
Mokhtari, Parisa D.
Courcelle, Charmain T.
Courcelle, Justin
author_facet Hamilton, Nicklas A.
Jehru, Avery E.
Samples, William N.
Wendel, Brian M.
Mokhtari, Parisa D.
Courcelle, Charmain T.
Courcelle, Justin
author_sort Hamilton, Nicklas A.
collection PubMed
description Accurately completing DNA replication when two forks converge is essential to genomic stability. The RecBCD helicase–nuclease complex plays a central role in completion by promoting resection and joining of the excess DNA created when replisomes converge. chi sequences alter RecBCD activity and localize with crossover hotspots during sexual events in bacteria, yet their functional role during chromosome replication remains unknown. Here, we use two-dimensional agarose gel analysis to show that chi induces replication on substrates containing convergent forks. The induced replication is processive but uncoupled with respect to leading and lagging strand synthesis and can be suppressed by ter sites which limit replisome progression. Our observations demonstrate that convergent replisomes create a substrate that is processed by RecBCD and that chi, when encountered, switches RecBCD from a degradative to replicative function. We propose that chi serves to functionally differentiate DNA ends created during completion, which require degradation, from those created by chromosomal double-strand breaks, which require resynthesis.
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spelling pubmed-100251582023-03-21 chi sequences switch the RecBCD helicase–nuclease complex from degradative to replicative modes during the completion of DNA replication Hamilton, Nicklas A. Jehru, Avery E. Samples, William N. Wendel, Brian M. Mokhtari, Parisa D. Courcelle, Charmain T. Courcelle, Justin J Biol Chem Research Article Accurately completing DNA replication when two forks converge is essential to genomic stability. The RecBCD helicase–nuclease complex plays a central role in completion by promoting resection and joining of the excess DNA created when replisomes converge. chi sequences alter RecBCD activity and localize with crossover hotspots during sexual events in bacteria, yet their functional role during chromosome replication remains unknown. Here, we use two-dimensional agarose gel analysis to show that chi induces replication on substrates containing convergent forks. The induced replication is processive but uncoupled with respect to leading and lagging strand synthesis and can be suppressed by ter sites which limit replisome progression. Our observations demonstrate that convergent replisomes create a substrate that is processed by RecBCD and that chi, when encountered, switches RecBCD from a degradative to replicative function. We propose that chi serves to functionally differentiate DNA ends created during completion, which require degradation, from those created by chromosomal double-strand breaks, which require resynthesis. American Society for Biochemistry and Molecular Biology 2023-02-11 /pmc/articles/PMC10025158/ /pubmed/36781123 http://dx.doi.org/10.1016/j.jbc.2023.103013 Text en © 2023 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Research Article
Hamilton, Nicklas A.
Jehru, Avery E.
Samples, William N.
Wendel, Brian M.
Mokhtari, Parisa D.
Courcelle, Charmain T.
Courcelle, Justin
chi sequences switch the RecBCD helicase–nuclease complex from degradative to replicative modes during the completion of DNA replication
title chi sequences switch the RecBCD helicase–nuclease complex from degradative to replicative modes during the completion of DNA replication
title_full chi sequences switch the RecBCD helicase–nuclease complex from degradative to replicative modes during the completion of DNA replication
title_fullStr chi sequences switch the RecBCD helicase–nuclease complex from degradative to replicative modes during the completion of DNA replication
title_full_unstemmed chi sequences switch the RecBCD helicase–nuclease complex from degradative to replicative modes during the completion of DNA replication
title_short chi sequences switch the RecBCD helicase–nuclease complex from degradative to replicative modes during the completion of DNA replication
title_sort chi sequences switch the recbcd helicase–nuclease complex from degradative to replicative modes during the completion of dna replication
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10025158/
https://www.ncbi.nlm.nih.gov/pubmed/36781123
http://dx.doi.org/10.1016/j.jbc.2023.103013
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