Characterization of all RND-type multidrug efflux transporters in Vibrio parahaemolyticus

Taira Matsuo, Koji Nakamura, Toshio Kodama, Taro Mikami, Hirotaka Hiyoshi, Tomofusa Tsuchiya, Wakano Ogawa, Teruo Kuroda

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13 Citations (Scopus)

Abstract

Resistance nodulation cell division (RND)-type efflux transporters play the main role in intrinsic resistance to various antimicrobial agents in many gram-negative bacteria. Here, we estimated 12 RND-type efflux transporter genes in Vibrio parahaemolyticus. Because VmeAB has already been characterized, we cloned the other 11 RND-type efflux transporter genes and characterized them in Escherichia coli KAM33 cells, a drug hypersusceptible strain. KAM33 expressing either VmeCD, VmeEF, or VmeYZ showed increased minimum inhibitory concentrations (MICs) for several antimicrobial agents. Additional four RND-type transporters were functional as efflux pumps only when co-expressed with VpoC, an outer membrane component in V. parahaemolyticus. Furthermore, VmeCD, VmeEF, and VmeYZ co-expressed with VpoC exhibited a broader substrate specificity and conferred higher resistance than that with TolC of E. coli. Deletion mutants of these transporter genes were constructed in V. parahaemolyticus. TM32 (ΔvmeAB and ΔvmeCD) had significantly decreased MICs for many antimicrobial agents and the number of viable cells after exposure to deoxycholate were markedly reduced. Strains in which 12 operons were all disrupted had very low MICs and much lower fluid accumulation in rabbit ileal loops. These results indicate that resistance nodulation cell division-type efflux transporters contribute not only to intrinsic resistance but also to exerting the virulence of V. parahaemolyticus. We characterized 12 RND-type multidrug efflux transporters in Vibrio parahaemolyticus. Deletion mutants of all twelve transporter genes showed higher sensitivity for several antibiotics and bile, and lower pathogenicity.

Original languageEnglish
Pages (from-to)725-742
Number of pages18
JournalMicrobiologyOpen
Volume2
Issue number5
DOIs
Publication statusPublished - Oct 1 2013

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Keywords

  • Drug resistance
  • Multidrug efflux transporter
  • RND
  • V. parahaemolyticus

ASJC Scopus subject areas

  • Microbiology

Cite this

Matsuo, T., Nakamura, K., Kodama, T., Mikami, T., Hiyoshi, H., Tsuchiya, T., Ogawa, W., & Kuroda, T. (2013). Characterization of all RND-type multidrug efflux transporters in Vibrio parahaemolyticus. MicrobiologyOpen, 2(5), 725-742. https://doi.org/10.1002/mbo3.100