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.
authors.affiliation: Department of Microbiology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Okayama, Japan.
authors.forename: Taira
authors.initials: T
authors.lastname: Matsuo
authors.forename: Koji
authors.initials: K
authors.lastname: Nakamura
authors.forename: Toshio
authors.initials: T
authors.lastname: Kodama
authors.forename: Taro
authors.initials: T
authors.lastname: Mikami
authors.forename: Hirotaka
authors.initials: H
authors.lastname: Hiyoshi
authors.forename: Tomofusa
authors.initials: T
authors.lastname: Tsuchiya
authors.forename: Wakano
authors.initials: W
authors.lastname: Ogawa
authors.forename: Teruo
authors.initials: T
authors.lastname: Kuroda
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000378:metabolism; D001426:Bacterial Proteins / Q000145:classification / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D017353:Gene Deletion; D015870:Gene Expression; D026901:Membrane Transport Proteins / Q000235:genetics / Q000378:metabolism; D008826:Microbial Sensitivity Tests; D050296:Microbial Viability / Q000187:drug effects; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000145:classification / Q000235:genetics* / Q000378:metabolism; D009876:Operon; D010802:Phylogeny; D011817:Rabbits; D019076:Transgenes; D014735:Vibrio Infections / Q000188:drug therapy / Q000382:microbiology; D014736:Vibrio parahaemolyticus / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism / Q000472:pathogenicity*; D014774:Virulence
chemical_list: D000900:Anti-Bacterial Agents; D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D026901:Membrane Transport Proteins; D027425:ATP-Binding Cassette, Sub-Family C Proteins; C073126:tolC protein, E coli
keywords: Drug resistance; RND; V. parahaemolyticus; multidrug efflux transporter
references.citation: J Infect Dis. 2000 May;181(5):1661-6
references.pmid: 10823766
references.citation: FEMS Microbiol Lett. 2002 Nov 19;217(1):57-63
references.pmid: 12445646
references.citation: Nat Rev Microbiol. 2006 Aug;4(8):629-36
references.pmid: 16845433
references.citation: Mol Microbiol. 2006 Jan;59(1):126-41
references.pmid: 16359323
references.citation: FEMS Microbiol Lett. 2001 Aug 7;202(1):139-43
references.pmid: 11506922
references.citation: Antimicrob Agents Chemother. 2003 Sep;47(9):3030-3
references.pmid: 12937021
references.citation: Mol Microbiol. 1996 Aug;21(4):713-24
references.pmid: 8878035
references.citation: FEMS Microbiol Lett. 1998 Aug 1;165(1):21-7
references.pmid: 9711836
references.citation: J Bacteriol. 2007 Nov;189(21):7600-9
references.pmid: 17720796
references.citation: J Bacteriol. 2005 Mar;187(6):1923-9
references.pmid: 15743938
references.citation: Microbiology (Reading). 2007 Dec;153(Pt 12):4129-4137
references.pmid: 18048926
references.citation: N Engl J Med. 2005 Oct 6;353(14):1463-70
references.pmid: 16207848
references.citation: J Bacteriol. 2001 Oct;183(20):5803-12
references.pmid: 11566977
references.citation: J Antimicrob Chemother. 2003 Oct;52(4):572-5
references.pmid: 12951344
references.citation: Antimicrob Agents Chemother. 1999 Dec;43(12):2975-83
references.pmid: 10582892
references.citation: J Bacteriol. 2003 Feb;185(4):1236-44
references.pmid: 12562793
references.citation: Microbiol Immunol. 2005;49(11):999-1002
references.pmid: 16301811
references.citation: J Bacteriol. 1996 Oct;178(20):5853-9
references.pmid: 8830678
references.citation: MMWR Morb Mortal Wkly Rep. 2006 Aug 11;55(31):854-6
references.pmid: 16902397
references.citation: Arch Intern Med. 1972 Oct;130(4):574-83
references.pmid: 4562152
references.citation: Antimicrob Agents Chemother. 1999 Feb;43(2):415-7
references.pmid: 9925549
references.citation: J Bacteriol. 2002 Sep;184(18):5036-44
references.pmid: 12193619
references.citation: Proc Natl Acad Sci U S A. 1999 Jun 22;96(13):7190-5
references.pmid: 10377390
references.citation: Infect Immun. 2001 Jul;69(7):4681-5
references.pmid: 11402016
references.citation: Hepatology. 1984 Sep-Oct;4(5 Suppl):4S-14S
references.pmid: 6384004
references.citation: Infect Immun. 2010 Apr;78(4):1772-80
references.pmid: 20086084
references.citation: J Biochem. 1994 Jun;115(6):1162-5
references.pmid: 7982898
references.citation: FEMS Microbiol Lett. 2003 Sep 26;226(2):281-4
references.pmid: 14553923
references.citation: FEMS Microbiol Rev. 2009 Mar;33(2):430-49
references.pmid: 19207745
references.citation: Emerg Infect Dis. 2005 Jan;11(1):129-31
references.pmid: 15705337
references.citation: Microbiol Mol Biol Rev. 2000 Dec;64(4):672-93
references.pmid: 11104814
references.citation: FEMS Microbiol Lett. 2006 Dec;265(1):76-80
references.pmid: 17107421
references.citation: J Biochem. 1994 May;115(5):868-74
references.pmid: 7961600
references.citation: Microbiology (Reading). 2002 Aug;148(Pt 8):2371-2381
references.pmid: 12177331
references.citation: Anal Biochem. 1987 Jan;160(1):47-56
references.pmid: 2952030
references.citation: Mol Microbiol. 1997 Jan;23(2):345-54
references.pmid: 9044268
references.citation: Biochem Biophys Res Commun. 1998 Mar 27;244(3):898-902
references.pmid: 9535764
references.citation: Biol Pharm Bull. 2008 Apr;31(4):577-82
references.pmid: 18379044
references.citation: J Infect Chemother. 2005 Jun;11(3):146-51
references.pmid: 15990979
references.citation: Microbiol Mol Biol Rev. 2002 Dec;66(4):671-701, table of contents
references.pmid: 12456787
references.citation: Arch Intern Med. 1999 Dec 13-27;159(22):2647-58
references.pmid: 10597755
references.citation: J Antimicrob Chemother. 2010 Jul;65(7):1372-6
references.pmid: 20495209
references.citation: Microbiol Immunol. 2003;47(12):937-43
references.pmid: 14695443
references.citation: J Clin Microbiol. 2000 Feb;38(2):578-85
references.pmid: 10655349
references.citation: Cell Microbiol. 2006 May;8(5):847-56
references.pmid: 16611233
references.citation: J Bacteriol. 1997 Apr;179(8):2512-8
references.pmid: 9098046
references.citation: J Bacteriol. 1993 Nov;175(22):7363-72
references.pmid: 8226684
references.citation: Lancet. 2003 Mar 1;361(9359):743-9
references.pmid: 12620739
references.citation: Microbiol Immunol. 2005;49(8):711-9
references.pmid: 16113500
references.citation: J Mol Biol. 1967 Jul 14;27(1):87-106
references.pmid: 6033613
references.citation: Antimicrob Agents Chemother. 2000 Mar;44(3):658-64
references.pmid: 10681335
references.citation: Antimicrob Agents Chemother. 2005 May;49(5):2133-6
references.pmid: 15855547
references.citation: PLoS One. 2010 Oct 13;5(10):e13365
references.pmid: 20967223
references.citation: Arch Microbiol. 2006 Sep;186(3):171-81
references.pmid: 16804679
references.citation: Int J Infect Dis. 2007 Mar;11(2):185-7
references.pmid: 16793305
references.citation: Kansenshogaku Zasshi. 2001 Jun;75(6):485-9
references.pmid: 11494566
references.citation: Antimicrob Agents Chemother. 2003 Sep;47(9):2990-2
references.pmid: 12937010
references.citation: J Bacteriol. 1967 Feb;93(2):642-8
references.pmid: 4289962
references.citation: Microbiol Immunol. 2007;51(11):1061-70
references.pmid: 18037783