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

Research output: Contribution to journalArticle

11 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 2013

Fingerprint

Vibrio parahaemolyticus
Cell Division
Microbial Sensitivity Tests
Anti-Infective Agents
Genes
Virulence
Escherichia coli
Deoxycholic Acid
Operon
Substrate Specificity
Gram-Negative Bacteria
Bile
Cell Count
Anti-Bacterial Agents
Rabbits
Membranes
Pharmaceutical Preparations

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., ... 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

Characterization of all RND-type multidrug efflux transporters in Vibrio parahaemolyticus. / Matsuo, Taira; Nakamura, Koji; Kodama, Toshio; Mikami, Taro; Hiyoshi, Hirotaka; Tsuchiya, Tomofusa; Ogawa, Wakano; Kuroda, Teruo.

In: MicrobiologyOpen, Vol. 2, No. 5, 10.2013, p. 725-742.

Research output: Contribution to journalArticle

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, vol. 2, no. 5, pp. 725-742. https://doi.org/10.1002/mbo3.100
Matsuo T, Nakamura K, Kodama T, Mikami T, Hiyoshi H, Tsuchiya T et al. Characterization of all RND-type multidrug efflux transporters in Vibrio parahaemolyticus. MicrobiologyOpen. 2013 Oct;2(5):725-742. https://doi.org/10.1002/mbo3.100
Matsuo, Taira ; Nakamura, Koji ; Kodama, Toshio ; Mikami, Taro ; Hiyoshi, Hirotaka ; Tsuchiya, Tomofusa ; Ogawa, Wakano ; Kuroda, Teruo. / Characterization of all RND-type multidrug efflux transporters in Vibrio parahaemolyticus. In: MicrobiologyOpen. 2013 ; Vol. 2, No. 5. pp. 725-742.
@article{d85f027a6bf94f87b896a1ae2478e86c,
title = "Characterization of all RND-type multidrug efflux transporters in Vibrio parahaemolyticus",
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.",
keywords = "Drug resistance, Multidrug efflux transporter, RND, V. parahaemolyticus",
author = "Taira Matsuo and Koji Nakamura and Toshio Kodama and Taro Mikami and Hirotaka Hiyoshi and Tomofusa Tsuchiya and Wakano Ogawa and Teruo Kuroda",
year = "2013",
month = "10",
doi = "10.1002/mbo3.100",
language = "English",
volume = "2",
pages = "725--742",
journal = "MicrobiologyOpen",
issn = "2045-8827",
publisher = "John Wiley and Sons Inc.",
number = "5",

}

TY - JOUR

T1 - Characterization of all RND-type multidrug efflux transporters in Vibrio parahaemolyticus

AU - Matsuo, Taira

AU - Nakamura, Koji

AU - Kodama, Toshio

AU - Mikami, Taro

AU - Hiyoshi, Hirotaka

AU - Tsuchiya, Tomofusa

AU - Ogawa, Wakano

AU - Kuroda, Teruo

PY - 2013/10

Y1 - 2013/10

N2 - 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.

AB - 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.

KW - Drug resistance

KW - Multidrug efflux transporter

KW - RND

KW - V. parahaemolyticus

UR - http://www.scopus.com/inward/record.url?scp=84885465600&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84885465600&partnerID=8YFLogxK

U2 - 10.1002/mbo3.100

DO - 10.1002/mbo3.100

M3 - Article

VL - 2

SP - 725

EP - 742

JO - MicrobiologyOpen

JF - MicrobiologyOpen

SN - 2045-8827

IS - 5

ER -