The Journal of antimicrobial chemotherapy · 2005
Characterization of the MacA-MacB efflux system in Neisseria gonorrhoeae.
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Rouquette-Loughlin Corinne E, Balthazar Jacqueline T, Shafer William M
Аннотация
OBJECTIVES: A homologue of the MacA-MacB ABC transporter of Escherichia coli, which recognizes and exports macrolides, was identified in Neisseria gonorrhoeae. This study was undertaken to determine whether gonococci could use the MacA-MacB homologue to express decreased susceptibility to macrolides.
METHODS: Techniques of DNA sequencing, gene cloning and expression of recombinant proteins in E. coli, gene mutation construction, transcriptional analysis and antimicrobial susceptibility testing were used in the study.
RESULTS: Although the gonococcal MacA-MacB efflux pump enhanced bacterial resistance to macrolides when overexpressed in an E. coli background, its loss in a gonococcal clinical isolate only slightly decreased bacterial resistance to azithromycin and erythromycin. However, a mutation in the -10 sequence of the promoter used in macAB expression enhanced the macrolide resistance of gonococci that produced a defective MtrC-MtrD-MtrE pump, which also recognizes macrolides.
CONCLUSIONS: The results from this study indicate that gonococci can employ both the MacA-MacB and MtrC-MtrD-MtrE efflux pumps to develop resistance to macrolides, particularly if mutations develop in the promoter that drives transcription of macAB.
METHODS: Techniques of DNA sequencing, gene cloning and expression of recombinant proteins in E. coli, gene mutation construction, transcriptional analysis and antimicrobial susceptibility testing were used in the study.
RESULTS: Although the gonococcal MacA-MacB efflux pump enhanced bacterial resistance to macrolides when overexpressed in an E. coli background, its loss in a gonococcal clinical isolate only slightly decreased bacterial resistance to azithromycin and erythromycin. However, a mutation in the -10 sequence of the promoter used in macAB expression enhanced the macrolide resistance of gonococci that produced a defective MtrC-MtrD-MtrE pump, which also recognizes macrolides.
CONCLUSIONS: The results from this study indicate that gonococci can employ both the MacA-MacB and MtrC-MtrD-MtrE efflux pumps to develop resistance to macrolides, particularly if mutations develop in the promoter that drives transcription of macAB.
Предметные рубрики MeSH
ATP-Binding Cassette TransportersAnti-Bacterial AgentsAzithromycinBacterial Outer Membrane ProteinsBacterial ProteinsBase SequenceCloning, MolecularDNA, BacterialDrug Resistance, BacterialErythromycinGene DeletionGene ExpressionLipoproteinsMacrolidesMembrane ProteinsMembrane Transport ProteinsMicrobial Sensitivity TestsMolecular Sequence DataMutationNeisseria gonorrhoeaeOpen Reading FramesOperonPromoter Regions, GeneticSequence Analysis, DNATranscription, Genetic