Phenotypic and Genome-Wide Analysis of an Antibiotic-Resistant Small Colony Variant

Small colony variants (SCVs) are slow-growing bacteria, which often show increased resistance to antibiotics and cause latent or recurrent infections. It is therefore important to understand the mechanisms at the basis of this phenotypic switch. One SCV (termed PAO-SCV) was isolated, showing high re...

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Veröffentlicht in:PloS one 2011-12, Vol.6 (12), p.e29276
Hauptverfasser: Wei, Qing, Tarighi, Saeed, Dötsch, Andreas, Häussler, Susanne, Müsken, Mathias, Wright, Victoria J, Cámara, Miguel, Williams, Paul, Haenen, Steven, Boerjan, Bart, Bogaerts, Annelies, Vierstraete, Evy, Verleyen, Peter, Schoofs, Liliane, Willaert, Ronnie, De Groote, Valérie N, Michiels, Jan, Vercammen, Ken, Crabbé, Aurélie, Cornelis, Pierre
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container_end_page
container_issue 12
container_start_page e29276
container_title PloS one
container_volume 6
creator Wei, Qing
Tarighi, Saeed
Dötsch, Andreas
Häussler, Susanne
Müsken, Mathias
Wright, Victoria J
Cámara, Miguel
Williams, Paul
Haenen, Steven
Boerjan, Bart
Bogaerts, Annelies
Vierstraete, Evy
Verleyen, Peter
Schoofs, Liliane
Willaert, Ronnie
De Groote, Valérie N
Michiels, Jan
Vercammen, Ken
Crabbé, Aurélie
Cornelis, Pierre
description Small colony variants (SCVs) are slow-growing bacteria, which often show increased resistance to antibiotics and cause latent or recurrent infections. It is therefore important to understand the mechanisms at the basis of this phenotypic switch. One SCV (termed PAO-SCV) was isolated, showing high resistance to gentamicin and to the cephalosporine cefotaxime. PAO-SCV was prone to reversion as evidenced by emergence of large colonies with a frequency of 10.sup.-5 on media without antibiotics while it was stably maintained in presence of gentamicin. PAO-SCV showed a delayed growth, defective motility, and strongly reduced levels of the quorum sensing Pseudomonas quinolone signal (PQS). Whole genome expression analysis further suggested a multi-layered antibiotic resistance mechanism, including simultaneous over-expression of two drug efflux pumps (MexAB-OprM, MexXY-OprM), the LPS modification operon arnBCADTEF, and the PhoP-PhoQ two-component system. Conversely, the genes for the synthesis of PQS were strongly down-regulated in PAO-SCV. Finally, genomic analysis revealed the presence of mutations in phoP and phoQ genes as well as in the mexZ gene encoding a repressor of the mexXY and mexAB-oprM genes. Only one mutation occurred only in REV, at nucleotide 1020 of the tufA gene, a paralog of tufB, both encoding the elongation factor Tu, causing a change of the rarely used aspartic acid codon GAU to the more common GAC, possibly causing an increase of tufA mRNA translation. High expression of phoP and phoQ was confirmed for the SCV variant while the revertant showed expression levels reduced to wild-type levels. By combining data coming from phenotypic, gene expression and proteome analysis, we could demonstrate that resistance to aminoglycosides in one SCV mutant is multifactorial including overexpression of efflux mechanisms, LPS modification and is accompanied by a drastic down-regulation of the Pseudomonas quinolone signal quorum sensing system.
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It is therefore important to understand the mechanisms at the basis of this phenotypic switch. One SCV (termed PAO-SCV) was isolated, showing high resistance to gentamicin and to the cephalosporine cefotaxime. PAO-SCV was prone to reversion as evidenced by emergence of large colonies with a frequency of 10.sup.-5 on media without antibiotics while it was stably maintained in presence of gentamicin. PAO-SCV showed a delayed growth, defective motility, and strongly reduced levels of the quorum sensing Pseudomonas quinolone signal (PQS). Whole genome expression analysis further suggested a multi-layered antibiotic resistance mechanism, including simultaneous over-expression of two drug efflux pumps (MexAB-OprM, MexXY-OprM), the LPS modification operon arnBCADTEF, and the PhoP-PhoQ two-component system. Conversely, the genes for the synthesis of PQS were strongly down-regulated in PAO-SCV. 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By combining data coming from phenotypic, gene expression and proteome analysis, we could demonstrate that resistance to aminoglycosides in one SCV mutant is multifactorial including overexpression of efflux mechanisms, LPS modification and is accompanied by a drastic down-regulation of the Pseudomonas quinolone signal quorum sensing system.</abstract><pub>Public Library of Science</pub><doi>10.1371/journal.pone.0029276</doi><tpages>e29276</tpages></addata></record>
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source DOAJ Directory of Open Access Journals; Public Library of Science (PLoS) Journals Open Access; EZB-FREE-00999 freely available EZB journals; PubMed Central; Free Full-Text Journals in Chemistry
subjects Aminoglycosides
Analysis
Aspartate
Genes
Genetic research
Genomes
Genomics
Health aspects
Infection
Microbial drug resistance
Translation (Genetics)
title Phenotypic and Genome-Wide Analysis of an Antibiotic-Resistant Small Colony Variant
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