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SmeOP-TolCSm efflux pump contributes to the multidrug resistance of Stenotrophomonas maltophilia.

ABSTRACT: A five-gene cluster, tolCSm-pcm-smeRo-smeO-smeP, of Stenotrophomonas maltophilia was characterized. The presence of smeOP and smeRo-pcm-tolCSm operons was verified by reverse transcription (RT)-PCR. Both operons were negatively regulated by the TetR-type transcriptional regulator SmeRo, as demonstrated by quantitative RT-PCR and a promoter-fusion assay. SmeO and SmeP were associated with TolCSm (the TolC protein of S. maltophilia) for the assembly of a resistance-nodulation-cell-division (RND)-type pump. The compounds extruded by SmeOP-TolCSm mainly included nalidixic acid, doxycycline, amikacin, gentamicin, erythromycin, leucomycin, carbonyl cyanide 3-chlorophenylhydrazone, crystal violet, sodium dodecyl sulfate, and tetrachlorosalicylanilide.

SUBMITTER: Lin CW

PROVIDER: S-EPMC4023765 | biostudies-literature | 2014

REPOSITORIES: biostudies-literature

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SmeOP-TolCSm efflux pump contributes to the multidrug resistance of Stenotrophomonas maltophilia.

Lin Cheng-Wen CW Huang Yi-Wei YW Hu Rouh-Mei RM Yang Tsuey-Ching TC

Antimicrobial agents and chemotherapy 20140106 4

A five-gene cluster, tolCSm-pcm-smeRo-smeO-smeP, of Stenotrophomonas maltophilia was characterized. The presence of smeOP and smeRo-pcm-tolCSm operons was verified by reverse transcription (RT)-PCR. Both operons were negatively regulated by the TetR-type transcriptional regulator SmeRo, as demonstrated by quantitative RT-PCR and a promoter-fusion assay. SmeO and SmeP were associated with TolCSm (the TolC protein of S. maltophilia) for the assembly of a resistance-nodulation-cell-division (RND)-t ...[more]

PMID: 24395237

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Project description:Clinical strains of Stenotrophomonas maltophilia are often highly resistant to multiple antibiotics, although the mechanisms of resistance are generally poorly understood. Multidrug resistant (MDR) strains were readily selected by plating a sensitive reference strain of the organism individually onto a variety of antibiotics, including tetracycline, chloramphenicol, ciprofloxacin, and norfloxacin. Tetracycline-selected MDR strains typically showed cross-resistance to erythromycin and fluoroquinolones and, in some instances, aminoglycosides. MDR mutants selected with the other agents generally displayed resistance to chloramphenicol and fluoroquinolones only, although two MDR strains (e.g., K1385) were also resistant to erythromycin and hypersusceptible to aminoglycosides. Many of the MDR strains expressed either moderate or high levels of a novel outer membrane protein (OMP) of ca. 50 kDa molecular mass, a phenotype typical of MDR strains of Pseudomonas aeruginosa hyperexpressing drug efflux systems. Indeed, the 50-kDa OMP of these S. maltophilia MDR strains reacted with antibody to OprM, the outer membrane component of the MexAB-OprM MDR efflux system of P. aeruginosa. Similarly, a ca. 110-kDa cytoplasmic membrane protein of these MDR strains also reacted with antibody to the MexB component of the P. aeruginosa pump. The outer and cytoplasmic membranes of several clinical S. maltophilia strains also reacted with the anti-OprM and anti-MexB antibodies. N-terminal amino acid sequencing of a cyanogen bromide-generated peptide of the 50-kDa OMP of MDR strain K1385, dubbed SmeM (Stenotrophomonas multidrug efflux), revealed it to be very similar to a number of outer membrane multidrug efflux components of P. aeruginosa and Pseudomonas putida. Deletion of the L1 and L2 beta-lactamase genes confirmed that these enzymes were responsible for the bulk of the beta-lactam resistance of K1385 and its parent. Still, overexpression of the MDR efflux mechanism in an L1- and L2-deficient derivative of K1385 did yield a modest increase in resistance to a few beta-lactams. These data are consistent with the MDR efflux mechanism(s) playing a role in the multidrug resistance of S. maltophilia.

Dataset Information

SmeOP-TolCSm efflux pump contributes to the multidrug resistance of Stenotrophomonas maltophilia.

Publications

SmeOP-TolCSm efflux pump contributes to the multidrug resistance of Stenotrophomonas maltophilia.

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