Biochemical characterization of the transcriptional regulator BzdR from Azoarcus sp. CIB.

12 páginas, 10 figuras, 1 tabla -- PAGS nros. 35694-35705

Detalles Bibliográficos
Autores: Durante-Rodríguez, Gonzalo, Valderrama, J. Andrés, Mancheño, Jose M., Rivas, Germán, Alfonso, Carlos, Arias-Palomo, Ernesto, Llorca, Óscar, García, José Luis, Díaz, Eduardo, Carmona, Manuel
Tipo de recurso: artículo
Fecha de publicación:2010
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/52529
Acceso en línea:http://hdl.handle.net/10261/52529
Access Level:acceso abierto
Palabra clave:Bacterial transcription
Gene regulation
Gene transcription
Repressor protein
Transcription repressor
Azoarcus
Bzd
RAnaerobic
Benzoates
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spelling Biochemical characterization of the transcriptional regulator BzdR from Azoarcus sp. CIB.Durante-Rodríguez, GonzaloValderrama, J. AndrésMancheño, Jose M.Rivas, GermánAlfonso, CarlosArias-Palomo, ErnestoLlorca, ÓscarGarcía, José LuisDíaz, EduardoCarmona, ManuelBacterial transcriptionGene regulationGene transcriptionRepressor proteinTranscription repressorAzoarcusBzdRAnaerobicBenzoates12 páginas, 10 figuras, 1 tabla -- PAGS nros. 35694-35705The BzdR transcriptional regulator that controls the P(N) promoter responsible for the anaerobic catabolism of benzoate in Azoarcus sp. CIB constitutes the prototype of a new subfamily of transcriptional regulators. Here, we provide some insights about the functional-structural relationships of the BzdR protein. Analytical ultracentrifugation studies revealed that BzdR is homodimeric in solution. An electron microscopy three-dimensional reconstruction of the BzdR dimer has been obtained, and the predicted structures of the respective N- and C-terminal domains of each BzdR monomer could be fitted into such a reconstruction. Gel retardation and ultracentrifugation experiments have shown that the binding of BzdR to its cognate promoter is cooperative. Different biochemical approaches revealed that the effector molecule benzoyl-CoA induces conformational changes in BzdR without affecting its oligomeric state. The BzdR-dependent inhibition of the P(N) promoter and its activation in the presence of benzoyl-CoA have been established by in vitro transcription assays. The monomeric BzdR4 and BzdR5 mutant regulators revealed that dimerization of BzdR is essential for DNA binding. Remarkably, a BzdRΔL protein lacking the linker region connecting the N- and C-terminal domains of BzdR is also dimeric and behaves as a super-repressor of the P(N) promoter. These data suggest that the linker region of BzdR is not essential for protein dimerization, but rather it is required to transfer the conformational changes induced by the benzoyl-CoA to the DNA binding domain leading to the release of the repressor. A model of action of the BzdR regulator has been proposed.This work was supported in part by Comisión Interministerial de Ciencia y Tecnología Grants BIO2006-05957, BIO2009-10438, BIO2008-04478-C03-03, BFU2007-67404/BMC, SAF2008-00451, and CSD2007-00005 and Comunidad Autónoma de Madrid Grants P-AMB-259-0505 and CAM S-BIO-0214-2006Peer reviewedAmerican Society for Biochemistry and Molecular Biology201220122010info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501http://hdl.handle.net/10261/52529reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Ingléshttp://dx.doi.org/ 10.1074/jbc.M110.143503info:eu-repo/semantics/openAccessoai:digital.csic.es:10261/525292026-05-22T06:33:51Z
dc.title.none.fl_str_mv Biochemical characterization of the transcriptional regulator BzdR from Azoarcus sp. CIB.
title Biochemical characterization of the transcriptional regulator BzdR from Azoarcus sp. CIB.
spellingShingle Biochemical characterization of the transcriptional regulator BzdR from Azoarcus sp. CIB.
Durante-Rodríguez, Gonzalo
Bacterial transcription
Gene regulation
Gene transcription
Repressor protein
Transcription repressor
Azoarcus
Bzd
RAnaerobic
Benzoates
title_short Biochemical characterization of the transcriptional regulator BzdR from Azoarcus sp. CIB.
title_full Biochemical characterization of the transcriptional regulator BzdR from Azoarcus sp. CIB.
title_fullStr Biochemical characterization of the transcriptional regulator BzdR from Azoarcus sp. CIB.
title_full_unstemmed Biochemical characterization of the transcriptional regulator BzdR from Azoarcus sp. CIB.
title_sort Biochemical characterization of the transcriptional regulator BzdR from Azoarcus sp. CIB.
dc.creator.none.fl_str_mv Durante-Rodríguez, Gonzalo
Valderrama, J. Andrés
Mancheño, Jose M.
Rivas, Germán
Alfonso, Carlos
Arias-Palomo, Ernesto
Llorca, Óscar
García, José Luis
Díaz, Eduardo
Carmona, Manuel
author Durante-Rodríguez, Gonzalo
author_facet Durante-Rodríguez, Gonzalo
Valderrama, J. Andrés
Mancheño, Jose M.
Rivas, Germán
Alfonso, Carlos
Arias-Palomo, Ernesto
Llorca, Óscar
García, José Luis
Díaz, Eduardo
Carmona, Manuel
author_role author
author2 Valderrama, J. Andrés
Mancheño, Jose M.
Rivas, Germán
Alfonso, Carlos
Arias-Palomo, Ernesto
Llorca, Óscar
García, José Luis
Díaz, Eduardo
Carmona, Manuel
author2_role author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Bacterial transcription
Gene regulation
Gene transcription
Repressor protein
Transcription repressor
Azoarcus
Bzd
RAnaerobic
Benzoates
topic Bacterial transcription
Gene regulation
Gene transcription
Repressor protein
Transcription repressor
Azoarcus
Bzd
RAnaerobic
Benzoates
description 12 páginas, 10 figuras, 1 tabla -- PAGS nros. 35694-35705
publishDate 2010
dc.date.none.fl_str_mv 2010
2012
2012
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/10261/52529
url http://hdl.handle.net/10261/52529
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv http://dx.doi.org/ 10.1074/jbc.M110.143503
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv American Society for Biochemistry and Molecular Biology
publisher.none.fl_str_mv American Society for Biochemistry and Molecular Biology
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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score 15.811543

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