Bacillus Subtilis RecA Interacts With and Loads RadA/Sms to Unwind Recombination Intermediates During Natural Chromosomal Transformation

During natural transformation Bacillus subtilis RecA, polymerized onto the incoming single-stranded (ss) DNA, catalyses DNA strand invasion resulting in a displacement loop (D-loop) intermediate. A null radA mutation impairs chromosomal transformation, and RadA/Sms unwinds forked DNA in the 5'→...

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Autores: Torres, Rubén, Serrano, Ester, Alonso, Juan Carlos
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2019
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/207545
Acceso en línea:http://hdl.handle.net/10261/207545
Access Level:acceso abierto
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spelling Bacillus Subtilis RecA Interacts With and Loads RadA/Sms to Unwind Recombination Intermediates During Natural Chromosomal TransformationTorres, RubénSerrano, EsterAlonso, Juan CarlosDuring natural transformation Bacillus subtilis RecA, polymerized onto the incoming single-stranded (ss) DNA, catalyses DNA strand invasion resulting in a displacement loop (D-loop) intermediate. A null radA mutation impairs chromosomal transformation, and RadA/Sms unwinds forked DNA in the 5'→3' direction. We show that in the absence of RadA/Sms competent cells require the RecG translocase for natural chromosomal transformation. RadA/Sms tetracysteine motif (C13A and C13R) variants, which fail to interact with RecA, are also deficient in plasmid transformation, but this defect is suppressed by inactivating recA. The RadA/Sms C13A and C13R variants bind ssDNA, and this interaction stimulates their ATPase activity. Wild-type (wt) RadA/Sms interacts with and inhibits the ATPase activity of RecA, but RadA/Sms C13A fails to do it. RadA/Sms and its variants, C13A and C13R, bound to the 5'-tail of a DNA substrate, unwind DNA in the 5'→3' direction. RecA interacts with and loads wt RadA/Sms to promote unwinding of a non-cognate 3'-tailed or 5'-fork DNA substrate, but RadA/Sms C13A or C13R fail to do it. We propose that wt RadA/Sms interaction with RecA is crucial to recruit the former onto D-loop DNA, and both proteins in concert catalyse D-loop extension to favour integration of ssDNA during chromosomal transformation.This work was supported in partby MINECO/FEDER BFU2015–67065-P and PGC2018–097054-B-I00 to J.C.APeer reviewedOxford University PressMinisterio de Ciencia, Innovación y Universidades (España)Alonso, Juan Carlos [0000-0002-5178-7179]Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]202020202019info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionhttp://hdl.handle.net/10261/207545reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Ingléshttps://doi.org/10.1093/nar/gkz647Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/2075452026-05-22T06:33:51Z
dc.title.none.fl_str_mv Bacillus Subtilis RecA Interacts With and Loads RadA/Sms to Unwind Recombination Intermediates During Natural Chromosomal Transformation
title Bacillus Subtilis RecA Interacts With and Loads RadA/Sms to Unwind Recombination Intermediates During Natural Chromosomal Transformation
spellingShingle Bacillus Subtilis RecA Interacts With and Loads RadA/Sms to Unwind Recombination Intermediates During Natural Chromosomal Transformation
Torres, Rubén
title_short Bacillus Subtilis RecA Interacts With and Loads RadA/Sms to Unwind Recombination Intermediates During Natural Chromosomal Transformation
title_full Bacillus Subtilis RecA Interacts With and Loads RadA/Sms to Unwind Recombination Intermediates During Natural Chromosomal Transformation
title_fullStr Bacillus Subtilis RecA Interacts With and Loads RadA/Sms to Unwind Recombination Intermediates During Natural Chromosomal Transformation
title_full_unstemmed Bacillus Subtilis RecA Interacts With and Loads RadA/Sms to Unwind Recombination Intermediates During Natural Chromosomal Transformation
title_sort Bacillus Subtilis RecA Interacts With and Loads RadA/Sms to Unwind Recombination Intermediates During Natural Chromosomal Transformation
dc.creator.none.fl_str_mv Torres, Rubén
Serrano, Ester
Alonso, Juan Carlos
author Torres, Rubén
author_facet Torres, Rubén
Serrano, Ester
Alonso, Juan Carlos
author_role author
author2 Serrano, Ester
Alonso, Juan Carlos
author2_role author
author
dc.contributor.none.fl_str_mv Ministerio de Ciencia, Innovación y Universidades (España)
Alonso, Juan Carlos [0000-0002-5178-7179]
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
description During natural transformation Bacillus subtilis RecA, polymerized onto the incoming single-stranded (ss) DNA, catalyses DNA strand invasion resulting in a displacement loop (D-loop) intermediate. A null radA mutation impairs chromosomal transformation, and RadA/Sms unwinds forked DNA in the 5'→3' direction. We show that in the absence of RadA/Sms competent cells require the RecG translocase for natural chromosomal transformation. RadA/Sms tetracysteine motif (C13A and C13R) variants, which fail to interact with RecA, are also deficient in plasmid transformation, but this defect is suppressed by inactivating recA. The RadA/Sms C13A and C13R variants bind ssDNA, and this interaction stimulates their ATPase activity. Wild-type (wt) RadA/Sms interacts with and inhibits the ATPase activity of RecA, but RadA/Sms C13A fails to do it. RadA/Sms and its variants, C13A and C13R, bound to the 5'-tail of a DNA substrate, unwind DNA in the 5'→3' direction. RecA interacts with and loads wt RadA/Sms to promote unwinding of a non-cognate 3'-tailed or 5'-fork DNA substrate, but RadA/Sms C13A or C13R fail to do it. We propose that wt RadA/Sms interaction with RecA is crucial to recruit the former onto D-loop DNA, and both proteins in concert catalyse D-loop extension to favour integration of ssDNA during chromosomal transformation.
publishDate 2019
dc.date.none.fl_str_mv 2019
2020
2020
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
Publisher's version
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/207545
url http://hdl.handle.net/10261/207545
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv https://doi.org/10.1093/nar/gkz647
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Oxford University Press
publisher.none.fl_str_mv Oxford University Press
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
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