Distribution of Holliday junctions and repair forks during Escherichia coli DNA double-strand break repair.

Accurate repair of DNA double-strand breaks (DSBs) is crucial for cell survival and genome integrity. In Escherichia coli, DSBs are repaired by homologous recombination (HR), using an undamaged sister chromosome as template. The DNA intermediates of this pathway are expected to be branched molecules...

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Autores principales:	Tahirah Yasmin, Benura Azeroglu, Charlotte A Cockram, David R F Leach
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2021
Materias:	Genetics QH426-470
Acceso en línea:	https://doaj.org/article/699560fd8ed641c9a2cef26281450fc3
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spelling	oai:doaj.org-article:699560fd8ed641c9a2cef26281450fc32021-12-02T20:02:51ZDistribution of Holliday junctions and repair forks during Escherichia coli DNA double-strand break repair.1553-73901553-740410.1371/journal.pgen.1009717https://doaj.org/article/699560fd8ed641c9a2cef26281450fc32021-08-01T00:00:00Zhttps://doi.org/10.1371/journal.pgen.1009717https://doaj.org/toc/1553-7390https://doaj.org/toc/1553-7404Accurate repair of DNA double-strand breaks (DSBs) is crucial for cell survival and genome integrity. In Escherichia coli, DSBs are repaired by homologous recombination (HR), using an undamaged sister chromosome as template. The DNA intermediates of this pathway are expected to be branched molecules that may include 4-way structures termed Holliday junctions (HJs), and 3-way structures such as D-loops and repair forks. Using a tool creating a site-specific, repairable DSB on only one of a pair of replicating sister chromosomes, we have determined how these branched DNA intermediates are distributed across a DNA region that is undergoing DSB repair. In cells, where branch migration and cleavage of HJs are limited by inactivation of the RuvABC complex, HJs and repair forks are principally accumulated within a distance of 12 kb from sites of recombination initiation, known as Chi, on each side of the engineered DSB. These branched DNA structures can even be detected in the region of DNA between the Chi sites flanking the DSB, a DNA segment not expected to be engaged in recombination initiation, and potentially degraded by RecBCD nuclease action. This is observed even in the absence of the branch migration and helicase activities of RuvAB, RadA, RecG, RecQ and PriA. The detection of full-length DNA fragments containing HJs in this central region implies that DSB repair can restore the two intact chromosomes, into which HJs can relocate prior to their resolution. The distribution of recombination intermediates across the 12kb region beyond Chi is altered in xonA, recJ and recQ mutants suggesting that, in the RecBCD pathway of DSB repair, exonuclease I stimulates the formation of repair forks and that RecJQ promotes strand-invasion at a distance from the recombination initiation sites.Tahirah YasminBenura AzerogluCharlotte A CockramDavid R F LeachPublic Library of Science (PLoS)articleGeneticsQH426-470ENPLoS Genetics, Vol 17, Iss 8, p e1009717 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Genetics QH426-470
spellingShingle	Genetics QH426-470 Tahirah Yasmin Benura Azeroglu Charlotte A Cockram David R F Leach Distribution of Holliday junctions and repair forks during Escherichia coli DNA double-strand break repair.
description	Accurate repair of DNA double-strand breaks (DSBs) is crucial for cell survival and genome integrity. In Escherichia coli, DSBs are repaired by homologous recombination (HR), using an undamaged sister chromosome as template. The DNA intermediates of this pathway are expected to be branched molecules that may include 4-way structures termed Holliday junctions (HJs), and 3-way structures such as D-loops and repair forks. Using a tool creating a site-specific, repairable DSB on only one of a pair of replicating sister chromosomes, we have determined how these branched DNA intermediates are distributed across a DNA region that is undergoing DSB repair. In cells, where branch migration and cleavage of HJs are limited by inactivation of the RuvABC complex, HJs and repair forks are principally accumulated within a distance of 12 kb from sites of recombination initiation, known as Chi, on each side of the engineered DSB. These branched DNA structures can even be detected in the region of DNA between the Chi sites flanking the DSB, a DNA segment not expected to be engaged in recombination initiation, and potentially degraded by RecBCD nuclease action. This is observed even in the absence of the branch migration and helicase activities of RuvAB, RadA, RecG, RecQ and PriA. The detection of full-length DNA fragments containing HJs in this central region implies that DSB repair can restore the two intact chromosomes, into which HJs can relocate prior to their resolution. The distribution of recombination intermediates across the 12kb region beyond Chi is altered in xonA, recJ and recQ mutants suggesting that, in the RecBCD pathway of DSB repair, exonuclease I stimulates the formation of repair forks and that RecJQ promotes strand-invasion at a distance from the recombination initiation sites.
format	article
author	Tahirah Yasmin Benura Azeroglu Charlotte A Cockram David R F Leach
author_facet	Tahirah Yasmin Benura Azeroglu Charlotte A Cockram David R F Leach
author_sort	Tahirah Yasmin
title	Distribution of Holliday junctions and repair forks during Escherichia coli DNA double-strand break repair.
title_short	Distribution of Holliday junctions and repair forks during Escherichia coli DNA double-strand break repair.
title_full	Distribution of Holliday junctions and repair forks during Escherichia coli DNA double-strand break repair.
title_fullStr	Distribution of Holliday junctions and repair forks during Escherichia coli DNA double-strand break repair.
title_full_unstemmed	Distribution of Holliday junctions and repair forks during Escherichia coli DNA double-strand break repair.
title_sort	distribution of holliday junctions and repair forks during escherichia coli dna double-strand break repair.
publisher	Public Library of Science (PLoS)
publishDate	2021
url	https://doaj.org/article/699560fd8ed641c9a2cef26281450fc3
work_keys_str_mv	AT tahirahyasmin distributionofhollidayjunctionsandrepairforksduringescherichiacolidnadoublestrandbreakrepair AT benuraazeroglu distributionofhollidayjunctionsandrepairforksduringescherichiacolidnadoublestrandbreakrepair AT charlotteacockram distributionofhollidayjunctionsandrepairforksduringescherichiacolidnadoublestrandbreakrepair AT davidrfleach distributionofhollidayjunctionsandrepairforksduringescherichiacolidnadoublestrandbreakrepair
_version_	1718375665699389440

Distribution of Holliday junctions and repair forks during Escherichia coli DNA double-strand break repair.

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