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Mechanisms of Resistance

Expression of Pseudomonas aeruginosa Multidrug Efflux Pumps MexA-MexB-OprM and MexC-MexD-OprJ in a Multidrug-Sensitive Escherichia coli Strain

Ramakrishnan Srikumar, Tatiana Kon, Naomasa Gotoh, Keith Poole
Ramakrishnan Srikumar
Department of Microbiology and Immunology, Queen’s University, Kingston, Ontario K7L 3N6, Canada, and
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Tatiana Kon
Department of Microbiology and Immunology, Queen’s University, Kingston, Ontario K7L 3N6, Canada, and
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Naomasa Gotoh
Department of Microbiology, Kyoto Pharmaceutical University, Yamashina, Kyoto 607, Japan
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Keith Poole
Department of Microbiology and Immunology, Queen’s University, Kingston, Ontario K7L 3N6, Canada, and
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DOI: 10.1128/AAC.42.1.65
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ABSTRACT

The mexCD-oprJ and mexAB-oprM operons encode components of two distinct multidrug efflux pumps inPseudomonas aeruginosa. To assess the contribution of individual components to antibiotic resistance and substrate specificity, these operons and their component genes were cloned and expressed in Escherichia coli. Western immunoblotting confirmed expression of the P. aeruginosa efflux pump components in E. coli strains expressing and deficient in the endogenous multidrug efflux system (AcrAB), although only the ΔacrAB strain, KZM120, demonstrated increased resistance to antibiotics in the presence of the P. aeruginosa efflux genes. E. coli KZM120 expressing MexAB-OprM showed increased resistance to quinolones, chloramphenicol, erythromycin, azithromycin, sodium dodecyl sulfate (SDS), crystal violet, novobiocin, and, significantly, several β-lactams, which is reminiscent of the operation of this pump in P. aeruginosa. This confirmed previous suggestions that MexAB-OprM provides a direct contribution to β-lactam resistance via the efflux of this group of antibiotics. An increase in antibiotic resistance, however, was not observed when MexAB or OprM alone was expressed in KZM120. Thus, despite the fact that β-lactams act within the periplasm, OprM alone is insufficient to provide resistance to these agents. E. coli KZM120 expressing MexCD-OprJ also showed increased resistance to quinolones, chloramphenicol, macrolides, SDS, and crystal violet, though not to most β-lactams or novobiocin, again somewhat reminiscent of the antibiotic resistance profile of MexCD-OprJ-expressing strains ofP. aeruginosa. Surprisingly, E. coli KZM120 expressing MexCD alone also showed an increase in resistance to these agents, while an OprJ-expressing KZM120 failed to demonstrate any increase in antibiotic resistance. MexCD-mediated resistance, however, was absent in a tolC mutant of KZM120, indicating that MexCD functions in KZM120 in conjunction with TolC, the previously identified outer membrane component of the AcrAB-TolC efflux system. These data confirm that a tripartite efflux pump is necessary for the efflux of all substrate antibiotics and that the P. aeruginosa multidrug efflux pumps are functional and retain their substrate specificity in E. coli.

  • Copyright © 1998 American Society for Microbiology
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Expression of Pseudomonas aeruginosa Multidrug Efflux Pumps MexA-MexB-OprM and MexC-MexD-OprJ in a Multidrug-Sensitive Escherichia coli Strain
Ramakrishnan Srikumar, Tatiana Kon, Naomasa Gotoh, Keith Poole
Antimicrobial Agents and Chemotherapy Jan 1998, 42 (1) 65-71; DOI: 10.1128/AAC.42.1.65

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Expression of Pseudomonas aeruginosa Multidrug Efflux Pumps MexA-MexB-OprM and MexC-MexD-OprJ in a Multidrug-Sensitive Escherichia coli Strain
Ramakrishnan Srikumar, Tatiana Kon, Naomasa Gotoh, Keith Poole
Antimicrobial Agents and Chemotherapy Jan 1998, 42 (1) 65-71; DOI: 10.1128/AAC.42.1.65
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KEYWORDS

Bacterial Proteins
Drug Resistance, Multiple
Escherichia coli
Pseudomonas aeruginosa

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