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Efflux pump genes of the resistance-nodulation-division family in Burkholderia cenocepacia genome

BACKGROUND: Burkholderia cenocepacia is recognized as opportunistic pathogen that can cause lung infections in cystic fibrosis patients. A hallmark of B. cenocepacia infections is the inability to eradicate the organism because of multiple intrinsic antibiotic resistance. As Resistance-Nodulation-Di...

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Autores principales: Guglierame, Paola, Pasca, Maria Rosalia, De Rossi, Edda, Buroni, Silvia, Arrigo, Patrizio, Manina, Giulia, Riccardi, Giovanna
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2006
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1557404/
https://www.ncbi.nlm.nih.gov/pubmed/16857052
http://dx.doi.org/10.1186/1471-2180-6-66
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author Guglierame, Paola
Pasca, Maria Rosalia
De Rossi, Edda
Buroni, Silvia
Arrigo, Patrizio
Manina, Giulia
Riccardi, Giovanna
author_facet Guglierame, Paola
Pasca, Maria Rosalia
De Rossi, Edda
Buroni, Silvia
Arrigo, Patrizio
Manina, Giulia
Riccardi, Giovanna
author_sort Guglierame, Paola
collection PubMed
description BACKGROUND: Burkholderia cenocepacia is recognized as opportunistic pathogen that can cause lung infections in cystic fibrosis patients. A hallmark of B. cenocepacia infections is the inability to eradicate the organism because of multiple intrinsic antibiotic resistance. As Resistance-Nodulation-Division (RND) efflux systems are responsible for much of the intrinsic multidrug resistance in Gram-negative bacteria, this study aims to identify RND genes in the B. cenocepacia genome and start to investigate their involvement into antimicrobial resistance. RESULTS: Genome analysis and homology searches revealed 14 open reading frames encoding putative drug efflux pumps belonging to RND family in B. cenocepacia J2315 strain. By reverse transcription (RT)-PCR analysis, it was found that orf3, orf9, orf11, and orf13 were expressed at detectable levels, while orf10 appeared to be weakly expressed in B. cenocepacia. Futhermore, orf3 was strongly induced by chloramphenicol. The orf2 conferred resistance to fluoroquinolones, tetraphenylphosphonium, streptomycin, and ethidium bromide when cloned and expressed in Escherichia coli KAM3, a strain lacking the multidrug efflux pump AcrAB. The orf2-overexpressing E. coli also accumulate low concentrations of ethidium bromide, which was restored to wild type level in the presence of CCCP, an energy uncoupler altering the energy of the drug efflux pump. CONCLUSION: The 14 RND pumps gene we have identified in the genome of B. cenocepacia suggest that active efflux could be a major mechanism underlying antimicrobial resistance in this microorganism. We have characterized the ORF2 pump, one of these 14 potential RND efflux systems. Its overexpression in E. coli conferred resistance to several antibiotics and to ethidium bromide but it remains to be determined if this pump play a significant role in the antimicrobial intrinsic resistance of B. cenocepacia. The characterization of antibiotic efflux pumps in B. cenocepacia is an obligatory step prior to the design of specific, potent bacterial inhibitors for the improved control of infectious diseases. Consequently, the topic deserves to be further investigated and future studies will involve systematic investigation on the function and expression of each of the RND efflux pump homologs.
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spelling pubmed-15574042006-08-30 Efflux pump genes of the resistance-nodulation-division family in Burkholderia cenocepacia genome Guglierame, Paola Pasca, Maria Rosalia De Rossi, Edda Buroni, Silvia Arrigo, Patrizio Manina, Giulia Riccardi, Giovanna BMC Microbiol Research Article BACKGROUND: Burkholderia cenocepacia is recognized as opportunistic pathogen that can cause lung infections in cystic fibrosis patients. A hallmark of B. cenocepacia infections is the inability to eradicate the organism because of multiple intrinsic antibiotic resistance. As Resistance-Nodulation-Division (RND) efflux systems are responsible for much of the intrinsic multidrug resistance in Gram-negative bacteria, this study aims to identify RND genes in the B. cenocepacia genome and start to investigate their involvement into antimicrobial resistance. RESULTS: Genome analysis and homology searches revealed 14 open reading frames encoding putative drug efflux pumps belonging to RND family in B. cenocepacia J2315 strain. By reverse transcription (RT)-PCR analysis, it was found that orf3, orf9, orf11, and orf13 were expressed at detectable levels, while orf10 appeared to be weakly expressed in B. cenocepacia. Futhermore, orf3 was strongly induced by chloramphenicol. The orf2 conferred resistance to fluoroquinolones, tetraphenylphosphonium, streptomycin, and ethidium bromide when cloned and expressed in Escherichia coli KAM3, a strain lacking the multidrug efflux pump AcrAB. The orf2-overexpressing E. coli also accumulate low concentrations of ethidium bromide, which was restored to wild type level in the presence of CCCP, an energy uncoupler altering the energy of the drug efflux pump. CONCLUSION: The 14 RND pumps gene we have identified in the genome of B. cenocepacia suggest that active efflux could be a major mechanism underlying antimicrobial resistance in this microorganism. We have characterized the ORF2 pump, one of these 14 potential RND efflux systems. Its overexpression in E. coli conferred resistance to several antibiotics and to ethidium bromide but it remains to be determined if this pump play a significant role in the antimicrobial intrinsic resistance of B. cenocepacia. The characterization of antibiotic efflux pumps in B. cenocepacia is an obligatory step prior to the design of specific, potent bacterial inhibitors for the improved control of infectious diseases. Consequently, the topic deserves to be further investigated and future studies will involve systematic investigation on the function and expression of each of the RND efflux pump homologs. BioMed Central 2006-07-20 /pmc/articles/PMC1557404/ /pubmed/16857052 http://dx.doi.org/10.1186/1471-2180-6-66 Text en Copyright © 2006 Guglierame et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( (http://creativecommons.org/licenses/by/2.0) ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Guglierame, Paola
Pasca, Maria Rosalia
De Rossi, Edda
Buroni, Silvia
Arrigo, Patrizio
Manina, Giulia
Riccardi, Giovanna
Efflux pump genes of the resistance-nodulation-division family in Burkholderia cenocepacia genome
title Efflux pump genes of the resistance-nodulation-division family in Burkholderia cenocepacia genome
title_full Efflux pump genes of the resistance-nodulation-division family in Burkholderia cenocepacia genome
title_fullStr Efflux pump genes of the resistance-nodulation-division family in Burkholderia cenocepacia genome
title_full_unstemmed Efflux pump genes of the resistance-nodulation-division family in Burkholderia cenocepacia genome
title_short Efflux pump genes of the resistance-nodulation-division family in Burkholderia cenocepacia genome
title_sort efflux pump genes of the resistance-nodulation-division family in burkholderia cenocepacia genome
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1557404/
https://www.ncbi.nlm.nih.gov/pubmed/16857052
http://dx.doi.org/10.1186/1471-2180-6-66
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