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Microbiology 155 (2009), 3509-3517; DOI  10.1099/mic.0.031260-0
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Microbiology 155 (2009), 3509-3517; DOI  10.1099/mic.0.031260-0
© 2009 Society for General Microbiology

Gene cloning and characteristics of the RND-type multidrug efflux pump MuxABC-OpmB possessing two RND components in Pseudomonas aeruginosa

Takehiko Mima1,{dagger},{ddagger}, Naoki Kohira1,{dagger}, Yang Li1, Hiroshi Sekiya1,§, Wakano Ogawa1, Teruo Kuroda1,2 and Tomofusa Tsuchiya1

1 Department of Molecular Microbiology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Tsushima, Kita-ku, Okayama 700-8530, Japan
2 Department of Genome Applied Microbiology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Tsushima, Kita-ku, Okayama 700-8530, Japan

muxA-muxB-muxC-opmB (formerly PA2528-PA2527-PA2526-opmB), encoding a putative resistance nodulation cell division (RND)-type multidrug efflux pump system, was cloned from Pseudomonas aeruginosa PAO1. Introduction of muxABC-opmB into P. aeruginosa YM64, a drug-hypersusceptible strain, led to elevated MICs of aztreonam, macrolides, novobiocin and tetracycline. Since muxB and muxC, both of which encode RND components, were essential for function, MuxABC-OpmB is thought to be a drug efflux pump with four components. One novobiocin-resistant mutant, PMX725, isolated from P. aeruginosa PMX7 showed elevated resistance not only to novobiocin but also to aztreonam, macrolides and tetracycline. Increased mRNA expression of muxABC-opmB was observed in the mutant PMX725 compared with the parental strain. Sequencing analysis revealed that a single-nucleotide insertion had occurred in the deduced promoter region for muxABC-opmB in PMX725. In this study, we have characterized the last RND-type multidrug efflux pump predicted from the genome sequence in P. aeruginosa.

Correspondence
Teruo Kuroda
tkuroda{at}cc.okayama-u.ac.jp

Abbreviations: RND, resistance nodulation cell division

{dagger}These authors contributed equally to this work.

{ddagger}Present address: Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, CO 80523-1682, USA.

§Present address: Department of Infectious Diseases, College of Pharmaceutical Sciences, Matsuyama University, Bunkyo-cho, Matsuyama, Ehime, 790-8578, Japan.






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