A marr-type regulator directly activates transcription from the Brucella abortus virB promoter by sharing a redundant role with HutC

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Type IV secretion systems (T4SS) are multiprotein structures that direct the translocation of specific molecules across the bacterial cell envelope. As in other bacteria, pathogenicity of the genus Brucella essentially depends on the integrity of the T4SS-encoding virB operon, whose expression is re...

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Autores principales:	Sieira, R., Arocena, G.M., Zorreguieta, A., Comerci, D.J., Ugalde, R.A.
Formato:	Artículo publishedVersion
Publicado:	2012
Materias:	deoxyribonuclease I protein hutc protein marR protein mdrA regulator protein unclassified drug VirB protein article binding site Brucella abortus DNA footprinting DNA sequence gel mobility shift assay gene expression regulation gene targeting nonhuman nucleotide sequence operon priority journal promoter region protein function transcription initiation transcription regulation type IV secretion system Binding Sites DNA Footprinting DNA, Bacterial Electrophoretic Mobility Shift Assay Gene Expression Regulation, Bacterial Promoter Regions, Genetic Protein Binding Transcription Factors Transcription, Genetic Virulence Factors Bacteria (microorganisms) Brucella Brucella melitensis biovar Abortus
Acceso en línea:	http://hdl.handle.net/20.500.12110/paper_00219193_v194_n23_p6431_Sieira http://repositoriouba.sisbi.uba.ar/gsdl/cgi-bin/library.cgi?a=d&c=artiaex&d=paper_00219193_v194_n23_p6431_Sieira_oai
Aporte de:	Repositorio Digital de la Universidad de Buenos Aires (UBA) de Universidad de Buenos Aires

id	I28-R145-paper_00219193_v194_n23_p6431_Sieira_oai
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spelling	I28-R145-paper_00219193_v194_n23_p6431_Sieira_oai2020-10-19 Sieira, R. Arocena, G.M. Zorreguieta, A. Comerci, D.J. Ugalde, R.A. 2012 Type IV secretion systems (T4SS) are multiprotein structures that direct the translocation of specific molecules across the bacterial cell envelope. As in other bacteria, pathogenicity of the genus Brucella essentially depends on the integrity of the T4SS-encoding virB operon, whose expression is regulated by multiple transcription factors belonging to different families. Previously, we identified IHF and HutC, two direct regulators of the virB genes that were isolated from total protein extracts of Brucella. Here, we report the identification of MdrA, a third regulatory element that was isolated using the same screening procedure. This transcription factor, which belongs to the MarR-family of transcriptional regulators, binds at two different sites of the virB promoter and regulates expression in a growth phase-dependent manner. Like other members of the MarR family, specific ligands were able to dissociate MdrA from DNA in vitro. Determination of the MdrA-binding sites by DNase I footprinting and analyses of protein-DNA complexes by electrophoresis mobility shift assays (EMSAs) showed that MdrA competes with IHF and HutC for the binding to the promoter because their target DNA sequences overlap. Unlike IHF, both MdrA and HutC bound to the promoter without inducing bending of DNA. Moreover, the two latter transcription factors activated virB expression to similar extents, and in doing so, they are functionally redundant. Taken together, our results show that MdrA is a regulatory element that directly modulates the activity of the virB promoter and is probably involved in coordinating gene expression in response to specific environmental signals. © 2012, American Society for Microbiology. Fil:Zorreguieta, A. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Ugalde, R.A. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. application/pdf http://hdl.handle.net/20.500.12110/paper_00219193_v194_n23_p6431_Sieira info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar J. Bacteriol. 2012;194(23):6431-6440 deoxyribonuclease I protein hutc protein marR protein mdrA regulator protein unclassified drug VirB protein article binding site Brucella abortus DNA footprinting DNA sequence gel mobility shift assay gene expression regulation gene targeting nonhuman nucleotide sequence operon priority journal promoter region protein function transcription initiation transcription regulation type IV secretion system Binding Sites Brucella abortus DNA Footprinting DNA, Bacterial Electrophoretic Mobility Shift Assay Gene Expression Regulation, Bacterial Promoter Regions, Genetic Protein Binding Transcription Factors Transcription, Genetic Virulence Factors Bacteria (microorganisms) Brucella Brucella melitensis biovar Abortus A marr-type regulator directly activates transcription from the Brucella abortus virB promoter by sharing a redundant role with HutC info:eu-repo/semantics/article info:ar-repo/semantics/artículo info:eu-repo/semantics/publishedVersion http://repositoriouba.sisbi.uba.ar/gsdl/cgi-bin/library.cgi?a=d&c=artiaex&d=paper_00219193_v194_n23_p6431_Sieira_oai
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-145
collection	Repositorio Digital de la Universidad de Buenos Aires (UBA)
topic	deoxyribonuclease I protein hutc protein marR protein mdrA regulator protein unclassified drug VirB protein article binding site Brucella abortus DNA footprinting DNA sequence gel mobility shift assay gene expression regulation gene targeting nonhuman nucleotide sequence operon priority journal promoter region protein function transcription initiation transcription regulation type IV secretion system Binding Sites Brucella abortus DNA Footprinting DNA, Bacterial Electrophoretic Mobility Shift Assay Gene Expression Regulation, Bacterial Promoter Regions, Genetic Protein Binding Transcription Factors Transcription, Genetic Virulence Factors Bacteria (microorganisms) Brucella Brucella melitensis biovar Abortus
spellingShingle	deoxyribonuclease I protein hutc protein marR protein mdrA regulator protein unclassified drug VirB protein article binding site Brucella abortus DNA footprinting DNA sequence gel mobility shift assay gene expression regulation gene targeting nonhuman nucleotide sequence operon priority journal promoter region protein function transcription initiation transcription regulation type IV secretion system Binding Sites Brucella abortus DNA Footprinting DNA, Bacterial Electrophoretic Mobility Shift Assay Gene Expression Regulation, Bacterial Promoter Regions, Genetic Protein Binding Transcription Factors Transcription, Genetic Virulence Factors Bacteria (microorganisms) Brucella Brucella melitensis biovar Abortus Sieira, R. Arocena, G.M. Zorreguieta, A. Comerci, D.J. Ugalde, R.A. A marr-type regulator directly activates transcription from the Brucella abortus virB promoter by sharing a redundant role with HutC
topic_facet	deoxyribonuclease I protein hutc protein marR protein mdrA regulator protein unclassified drug VirB protein article binding site Brucella abortus DNA footprinting DNA sequence gel mobility shift assay gene expression regulation gene targeting nonhuman nucleotide sequence operon priority journal promoter region protein function transcription initiation transcription regulation type IV secretion system Binding Sites Brucella abortus DNA Footprinting DNA, Bacterial Electrophoretic Mobility Shift Assay Gene Expression Regulation, Bacterial Promoter Regions, Genetic Protein Binding Transcription Factors Transcription, Genetic Virulence Factors Bacteria (microorganisms) Brucella Brucella melitensis biovar Abortus
description	Type IV secretion systems (T4SS) are multiprotein structures that direct the translocation of specific molecules across the bacterial cell envelope. As in other bacteria, pathogenicity of the genus Brucella essentially depends on the integrity of the T4SS-encoding virB operon, whose expression is regulated by multiple transcription factors belonging to different families. Previously, we identified IHF and HutC, two direct regulators of the virB genes that were isolated from total protein extracts of Brucella. Here, we report the identification of MdrA, a third regulatory element that was isolated using the same screening procedure. This transcription factor, which belongs to the MarR-family of transcriptional regulators, binds at two different sites of the virB promoter and regulates expression in a growth phase-dependent manner. Like other members of the MarR family, specific ligands were able to dissociate MdrA from DNA in vitro. Determination of the MdrA-binding sites by DNase I footprinting and analyses of protein-DNA complexes by electrophoresis mobility shift assays (EMSAs) showed that MdrA competes with IHF and HutC for the binding to the promoter because their target DNA sequences overlap. Unlike IHF, both MdrA and HutC bound to the promoter without inducing bending of DNA. Moreover, the two latter transcription factors activated virB expression to similar extents, and in doing so, they are functionally redundant. Taken together, our results show that MdrA is a regulatory element that directly modulates the activity of the virB promoter and is probably involved in coordinating gene expression in response to specific environmental signals. © 2012, American Society for Microbiology.
format	Artículo Artículo publishedVersion
author	Sieira, R. Arocena, G.M. Zorreguieta, A. Comerci, D.J. Ugalde, R.A.
author_facet	Sieira, R. Arocena, G.M. Zorreguieta, A. Comerci, D.J. Ugalde, R.A.
author_sort	Sieira, R.
title	A marr-type regulator directly activates transcription from the Brucella abortus virB promoter by sharing a redundant role with HutC
title_short	A marr-type regulator directly activates transcription from the Brucella abortus virB promoter by sharing a redundant role with HutC
title_full	A marr-type regulator directly activates transcription from the Brucella abortus virB promoter by sharing a redundant role with HutC
title_fullStr	A marr-type regulator directly activates transcription from the Brucella abortus virB promoter by sharing a redundant role with HutC
title_full_unstemmed	A marr-type regulator directly activates transcription from the Brucella abortus virB promoter by sharing a redundant role with HutC
title_sort	marr-type regulator directly activates transcription from the brucella abortus virb promoter by sharing a redundant role with hutc
publishDate	2012
url	http://hdl.handle.net/20.500.12110/paper_00219193_v194_n23_p6431_Sieira http://repositoriouba.sisbi.uba.ar/gsdl/cgi-bin/library.cgi?a=d&c=artiaex&d=paper_00219193_v194_n23_p6431_Sieira_oai
work_keys_str_mv	AT sieirar amarrtyperegulatordirectlyactivatestranscriptionfromthebrucellaabortusvirbpromoterbysharingaredundantrolewithhutc AT arocenagm amarrtyperegulatordirectlyactivatestranscriptionfromthebrucellaabortusvirbpromoterbysharingaredundantrolewithhutc AT zorreguietaa amarrtyperegulatordirectlyactivatestranscriptionfromthebrucellaabortusvirbpromoterbysharingaredundantrolewithhutc AT comercidj amarrtyperegulatordirectlyactivatestranscriptionfromthebrucellaabortusvirbpromoterbysharingaredundantrolewithhutc AT ugaldera amarrtyperegulatordirectlyactivatestranscriptionfromthebrucellaabortusvirbpromoterbysharingaredundantrolewithhutc AT sieirar marrtyperegulatordirectlyactivatestranscriptionfromthebrucellaabortusvirbpromoterbysharingaredundantrolewithhutc AT arocenagm marrtyperegulatordirectlyactivatestranscriptionfromthebrucellaabortusvirbpromoterbysharingaredundantrolewithhutc AT zorreguietaa marrtyperegulatordirectlyactivatestranscriptionfromthebrucellaabortusvirbpromoterbysharingaredundantrolewithhutc AT comercidj marrtyperegulatordirectlyactivatestranscriptionfromthebrucellaabortusvirbpromoterbysharingaredundantrolewithhutc AT ugaldera marrtyperegulatordirectlyactivatestranscriptionfromthebrucellaabortusvirbpromoterbysharingaredundantrolewithhutc
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A marr-type regulator directly activates transcription from the Brucella abortus virB promoter by sharing a redundant role with HutC

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