Interplay between two RND systems mediating antimicrobial resistance in brucella suis

The RND-type efflux pumps are responsible for the multidrug resistance phenotype observed in many clinically relevant species. Also, RND pumps have been implicated in physiological processes, with roles in the virulence mechanisms of several pathogenic bacteria. We have previously shown that the Bep...

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Autores principales:	Martín, Fernando Ariel, Posadas, Diana M., Zorreguieta, Angeles
Publicado:	2009
Materias:	ampicillin carrier protein ciprofloxacin crystal violet deoxycholic acid doxycycline ethidium bromide norfloxacin tetracycline antiinfective agent bacterial protein ethidium repressor protein antibiotic resistance article Brucella suis controlled study gene repression nonhuman phenotype priority journal promoter region amino acid sequence drug effect epithelium cell gene deletion gene dosage gene expression regulation genetics HeLa cell human metabolism microbiological examination microbiology pathogenicity physiology sequence alignment virulence Brucella melitensis biovar Suis Amino Acid Sequence Anti-Bacterial Agents Bacterial Proteins Deoxycholic Acid Drug Resistance, Bacterial Epithelial Cells Ethidium Gene Deletion Gene Dosage Gene Expression Regulation, Bacterial Gentian Violet Hela Cells Humans Membrane Transport Proteins Microbial Sensitivity Tests Repressor Proteins Sequence Alignment Virulence
Acceso en línea:	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00219193_v191_n8_p2530_Martin http://hdl.handle.net/20.500.12110/paper_00219193_v191_n8_p2530_Martin
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

id	paper:paper_00219193_v191_n8_p2530_Martin
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spelling	paper:paper_00219193_v191_n8_p2530_Martin2023-06-08T14:43:11Z Interplay between two RND systems mediating antimicrobial resistance in brucella suis Martín, Fernando Ariel Posadas, Diana M. Zorreguieta, Angeles ampicillin carrier protein ciprofloxacin crystal violet deoxycholic acid doxycycline ethidium bromide norfloxacin tetracycline antiinfective agent bacterial protein carrier protein crystal violet deoxycholic acid ethidium repressor protein antibiotic resistance article Brucella suis controlled study gene repression nonhuman phenotype priority journal promoter region amino acid sequence drug effect epithelium cell gene deletion gene dosage gene expression regulation genetics HeLa cell human metabolism microbiological examination microbiology pathogenicity physiology sequence alignment virulence Brucella melitensis biovar Suis Amino Acid Sequence Anti-Bacterial Agents Bacterial Proteins Brucella suis Deoxycholic Acid Drug Resistance, Bacterial Epithelial Cells Ethidium Gene Deletion Gene Dosage Gene Expression Regulation, Bacterial Gentian Violet Hela Cells Humans Membrane Transport Proteins Microbial Sensitivity Tests Repressor Proteins Sequence Alignment Virulence The RND-type efflux pumps are responsible for the multidrug resistance phenotype observed in many clinically relevant species. Also, RND pumps have been implicated in physiological processes, with roles in the virulence mechanisms of several pathogenic bacteria. We have previously shown that the BepC outer membrane factor of Brucella suis is involved in the efflux of diverse drugs, probably as part of a tripartite complex with an inner membrane translocase. In the present work, we characterize two membrane fusion protein-RND translocases of B. suis encoded by the bepDE and bepFG loci. MIC assays showed that the B. suis AbepE mutant was more sensitive to deoxycholate (DOC), ethidium bromide, and crystal violet. Furthermore, multicopy bepDE increased resistance to DOC and crystal violet and also to other drugs, including ampicillin, norfloxacin, ciprofloxacin, tetracycline, and doxycycline. In contrast to the ΔbepE mutant, the resistance profile of B. suis remained unaltered when the other RND gene (bepG) was deleted. However, the ΔbepE ΔbepG double mutant showed a more severe phenotype than the ΔbepE mutant, indicating that BepFG also contributes to drug resistance. An open reading frame (bepR) coding for a putative regulatory protein of the TetR family was found upstream of the bepDE locus. BepR strongly repressed the activity of the bepDE promoter, but DOC released the repression mediated by BepR. A clear induction of the bepFG promoter activity was observed only in the BepDE-defective mutant, indicating a regulatory interplay between the two RND efflux pumps. Although only the BepFG-defective mutant showed a moderate attenuation in model cells, the activities of both bepDE and bepFG promoters were induced in the intracellular environment of HeLa cells. Our results show that B. suis harbors two functional RND efflux pumps that may contribute to virulence. Copyright © 2009, American Society for Microbiology. All Rights Reserved. Fil:Martin, F.A. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Posadas, D.M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Zorreguieta, A. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 2009 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00219193_v191_n8_p2530_Martin http://hdl.handle.net/20.500.12110/paper_00219193_v191_n8_p2530_Martin
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	ampicillin carrier protein ciprofloxacin crystal violet deoxycholic acid doxycycline ethidium bromide norfloxacin tetracycline antiinfective agent bacterial protein carrier protein crystal violet deoxycholic acid ethidium repressor protein antibiotic resistance article Brucella suis controlled study gene repression nonhuman phenotype priority journal promoter region amino acid sequence drug effect epithelium cell gene deletion gene dosage gene expression regulation genetics HeLa cell human metabolism microbiological examination microbiology pathogenicity physiology sequence alignment virulence Brucella melitensis biovar Suis Amino Acid Sequence Anti-Bacterial Agents Bacterial Proteins Brucella suis Deoxycholic Acid Drug Resistance, Bacterial Epithelial Cells Ethidium Gene Deletion Gene Dosage Gene Expression Regulation, Bacterial Gentian Violet Hela Cells Humans Membrane Transport Proteins Microbial Sensitivity Tests Repressor Proteins Sequence Alignment Virulence
spellingShingle	ampicillin carrier protein ciprofloxacin crystal violet deoxycholic acid doxycycline ethidium bromide norfloxacin tetracycline antiinfective agent bacterial protein carrier protein crystal violet deoxycholic acid ethidium repressor protein antibiotic resistance article Brucella suis controlled study gene repression nonhuman phenotype priority journal promoter region amino acid sequence drug effect epithelium cell gene deletion gene dosage gene expression regulation genetics HeLa cell human metabolism microbiological examination microbiology pathogenicity physiology sequence alignment virulence Brucella melitensis biovar Suis Amino Acid Sequence Anti-Bacterial Agents Bacterial Proteins Brucella suis Deoxycholic Acid Drug Resistance, Bacterial Epithelial Cells Ethidium Gene Deletion Gene Dosage Gene Expression Regulation, Bacterial Gentian Violet Hela Cells Humans Membrane Transport Proteins Microbial Sensitivity Tests Repressor Proteins Sequence Alignment Virulence Martín, Fernando Ariel Posadas, Diana M. Zorreguieta, Angeles Interplay between two RND systems mediating antimicrobial resistance in brucella suis
topic_facet	ampicillin carrier protein ciprofloxacin crystal violet deoxycholic acid doxycycline ethidium bromide norfloxacin tetracycline antiinfective agent bacterial protein carrier protein crystal violet deoxycholic acid ethidium repressor protein antibiotic resistance article Brucella suis controlled study gene repression nonhuman phenotype priority journal promoter region amino acid sequence drug effect epithelium cell gene deletion gene dosage gene expression regulation genetics HeLa cell human metabolism microbiological examination microbiology pathogenicity physiology sequence alignment virulence Brucella melitensis biovar Suis Amino Acid Sequence Anti-Bacterial Agents Bacterial Proteins Brucella suis Deoxycholic Acid Drug Resistance, Bacterial Epithelial Cells Ethidium Gene Deletion Gene Dosage Gene Expression Regulation, Bacterial Gentian Violet Hela Cells Humans Membrane Transport Proteins Microbial Sensitivity Tests Repressor Proteins Sequence Alignment Virulence
description	The RND-type efflux pumps are responsible for the multidrug resistance phenotype observed in many clinically relevant species. Also, RND pumps have been implicated in physiological processes, with roles in the virulence mechanisms of several pathogenic bacteria. We have previously shown that the BepC outer membrane factor of Brucella suis is involved in the efflux of diverse drugs, probably as part of a tripartite complex with an inner membrane translocase. In the present work, we characterize two membrane fusion protein-RND translocases of B. suis encoded by the bepDE and bepFG loci. MIC assays showed that the B. suis AbepE mutant was more sensitive to deoxycholate (DOC), ethidium bromide, and crystal violet. Furthermore, multicopy bepDE increased resistance to DOC and crystal violet and also to other drugs, including ampicillin, norfloxacin, ciprofloxacin, tetracycline, and doxycycline. In contrast to the ΔbepE mutant, the resistance profile of B. suis remained unaltered when the other RND gene (bepG) was deleted. However, the ΔbepE ΔbepG double mutant showed a more severe phenotype than the ΔbepE mutant, indicating that BepFG also contributes to drug resistance. An open reading frame (bepR) coding for a putative regulatory protein of the TetR family was found upstream of the bepDE locus. BepR strongly repressed the activity of the bepDE promoter, but DOC released the repression mediated by BepR. A clear induction of the bepFG promoter activity was observed only in the BepDE-defective mutant, indicating a regulatory interplay between the two RND efflux pumps. Although only the BepFG-defective mutant showed a moderate attenuation in model cells, the activities of both bepDE and bepFG promoters were induced in the intracellular environment of HeLa cells. Our results show that B. suis harbors two functional RND efflux pumps that may contribute to virulence. Copyright © 2009, American Society for Microbiology. All Rights Reserved.
author	Martín, Fernando Ariel Posadas, Diana M. Zorreguieta, Angeles
author_facet	Martín, Fernando Ariel Posadas, Diana M. Zorreguieta, Angeles
author_sort	Martín, Fernando Ariel
title	Interplay between two RND systems mediating antimicrobial resistance in brucella suis
title_short	Interplay between two RND systems mediating antimicrobial resistance in brucella suis
title_full	Interplay between two RND systems mediating antimicrobial resistance in brucella suis
title_fullStr	Interplay between two RND systems mediating antimicrobial resistance in brucella suis
title_full_unstemmed	Interplay between two RND systems mediating antimicrobial resistance in brucella suis
title_sort	interplay between two rnd systems mediating antimicrobial resistance in brucella suis
publishDate	2009
url	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00219193_v191_n8_p2530_Martin http://hdl.handle.net/20.500.12110/paper_00219193_v191_n8_p2530_Martin
work_keys_str_mv	AT martinfernandoariel interplaybetweentworndsystemsmediatingantimicrobialresistanceinbrucellasuis AT posadasdianam interplaybetweentworndsystemsmediatingantimicrobialresistanceinbrucellasuis AT zorreguietaangeles interplaybetweentworndsystemsmediatingantimicrobialresistanceinbrucellasuis
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Interplay between two RND systems mediating antimicrobial resistance in brucella suis

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