Temperature-dependent hypermutational phenotype in recA mutants of Thermus thermophilus HB27

The recA gene from Thermus thermophilus HB27 was cloned and engineered to obtain insertion (recA::kat) and deletion (delta-recA) derivatives. Transcription of recA in this extreme thermophile was induced by mitomycin C, leading to the synthesis of a monocistronic mRNA. This DNA damage-mediated induc...

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Autores: Castán, Pablo, Casares, Lorena, Barbé, Jordi, Berenguer, José
Formato: artículo
Fecha de publicación:2003
País:España
Recursos:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/8289
Acesso em linha:http://hdl.handle.net/10261/8289
Access Level:acceso abierto
Palavra-chave:Thermus thermophilus
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spelling Temperature-dependent hypermutational phenotype in recA mutants of Thermus thermophilus HB27Castán, PabloCasares, LorenaBarbé, JordiBerenguer, JoséThermus thermophilusThe recA gene from Thermus thermophilus HB27 was cloned and engineered to obtain insertion (recA::kat) and deletion (delta-recA) derivatives. Transcription of recA in this extreme thermophile was induced by mitomycin C, leading to the synthesis of a monocistronic mRNA. This DNA damage-mediated induction was dependent on the integrity of recA. In addition to UV sensitivity, the recA mutants of T. thermophilus showed severe pleiotropic defects, ranging from irregular nucleoid condensation and segregation to a dramatic reduction in viability during culture. An increase in the frequency of both carotenoidless and auxotrophic mutants within surviving cells of the delta-recA strain indicated a high mutation rate. As RecA is not required for plasmid transformation, we have used the alpha-lacZ gene fragment and the ampicillin resistance gene from Escherichia coli as passenger reporters to confirm such high mutation rates. Our data support the idea that the absence of RecA results in a hypermutational phenotype in T. thermophilus. Furthermore, a direct relationship is deduced between the growth temperature and mutation rate, which finally has a deleterious effect on cell survival in the absence of RecA.This work was supported by grants BIO2001-1267 to J. Berenguer and BMC2001-2065 and 2001SGR-206 to J. Barbé and by an institutional grant from Fundación Ramón Areces to CBMSO. Pablo Castán held a fellowship from the Ministerio de Educación y Cultura.Peer reviewedAmerican Society for MicrobiologyFundación Ramón ArecesMinisterio de Educación y Cultura (España)200820082003info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501633090 bytesapplication/pdfhttp://hdl.handle.net/10261/8289reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Ingléshttp://dx.doi.org/10.1128/JB.185.16.4901-4907.2003info:eu-repo/semantics/openAccessoai:digital.csic.es:10261/82892026-05-22T06:33:51Z
dc.title.none.fl_str_mv Temperature-dependent hypermutational phenotype in recA mutants of Thermus thermophilus HB27
title Temperature-dependent hypermutational phenotype in recA mutants of Thermus thermophilus HB27
spellingShingle Temperature-dependent hypermutational phenotype in recA mutants of Thermus thermophilus HB27
Castán, Pablo
Thermus thermophilus
title_short Temperature-dependent hypermutational phenotype in recA mutants of Thermus thermophilus HB27
title_full Temperature-dependent hypermutational phenotype in recA mutants of Thermus thermophilus HB27
title_fullStr Temperature-dependent hypermutational phenotype in recA mutants of Thermus thermophilus HB27
title_full_unstemmed Temperature-dependent hypermutational phenotype in recA mutants of Thermus thermophilus HB27
title_sort Temperature-dependent hypermutational phenotype in recA mutants of Thermus thermophilus HB27
dc.creator.none.fl_str_mv Castán, Pablo
Casares, Lorena
Barbé, Jordi
Berenguer, José
author Castán, Pablo
author_facet Castán, Pablo
Casares, Lorena
Barbé, Jordi
Berenguer, José
author_role author
author2 Casares, Lorena
Barbé, Jordi
Berenguer, José
author2_role author
author
author
dc.contributor.none.fl_str_mv Fundación Ramón Areces
Ministerio de Educación y Cultura (España)
dc.subject.none.fl_str_mv Thermus thermophilus
topic Thermus thermophilus
description The recA gene from Thermus thermophilus HB27 was cloned and engineered to obtain insertion (recA::kat) and deletion (delta-recA) derivatives. Transcription of recA in this extreme thermophile was induced by mitomycin C, leading to the synthesis of a monocistronic mRNA. This DNA damage-mediated induction was dependent on the integrity of recA. In addition to UV sensitivity, the recA mutants of T. thermophilus showed severe pleiotropic defects, ranging from irregular nucleoid condensation and segregation to a dramatic reduction in viability during culture. An increase in the frequency of both carotenoidless and auxotrophic mutants within surviving cells of the delta-recA strain indicated a high mutation rate. As RecA is not required for plasmid transformation, we have used the alpha-lacZ gene fragment and the ampicillin resistance gene from Escherichia coli as passenger reporters to confirm such high mutation rates. Our data support the idea that the absence of RecA results in a hypermutational phenotype in T. thermophilus. Furthermore, a direct relationship is deduced between the growth temperature and mutation rate, which finally has a deleterious effect on cell survival in the absence of RecA.
publishDate 2003
dc.date.none.fl_str_mv 2003
2008
2008
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
format article
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/8289
url http://hdl.handle.net/10261/8289
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv http://dx.doi.org/10.1128/JB.185.16.4901-4907.2003
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 633090 bytes
application/pdf
dc.publisher.none.fl_str_mv American Society for Microbiology
publisher.none.fl_str_mv American Society for Microbiology
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
repository.name.fl_str_mv
repository.mail.fl_str_mv
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