1887

Abstract

Genes and , encoding a multidrug efflux transporter in the halophilic bacterium , have been cloned using a drug-hypersusceptible strain as the host. Cells of KAM33 (Δ Δ) carrying the region from conferred much higher MICs for a variety of antimicrobial agents than did control cells. Cells possessing VmeAB under energized conditions maintained very low intracellular concentrations of ethidium. This was as expected for an energy-dependent efflux system, and supports the notion – based on sequence homology – that VmeAB belongs to the resistance nodulation cell division (RND) family of multidrug efflux transporters. It is likely that VmeAB forms functional complexes with the outer-membrane protein TolC in , because introduction of into cells of KAM43, which lacks the gene, failed to elevate the MICs for any of the antimicrobial agents tested. Therefore, a homologue of was also cloned, designated , and was introduced together with into cells of KAM43. The MICs of all agents tested were raised and were comparable to the values observed in KAM33 harbouring a plasmid carrying . Finally, a -deficient mutant of was constructed (designated TM3). TM3 showed slightly higher susceptibility than the parental to some antimicrobial agents. Survival rate of the TM3 when exposed to deoxycholate decreased compared with that of the parent.

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2007-12-01
2020-08-10
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References

  1. Baumann P., Schubert R. H. W.. 1984; Family II. Vibrionaceae . In Bergey's Manual of Systematic Bacteriology vol. 1 pp516–550 Edited by Krieg N. R., Holt J. G. Baltimore: Williams & Wilkins;
  2. Berns K. I., Thomas C. A. Jr. 1965; Isolation of high molecular weight DNA from Haemophilus influenzae . J Mol Biol11:476–490
    [Google Scholar]
  3. Bina J. E., Mekalanos J. J.. 2001; Vibrio cholerae tolC is required for bile resistance and colonization. Infect Immun69:4681–4685
    [Google Scholar]
  4. Bina J. E., Provenzano D., Wang C., Bina X. R., Mekalanos J. J.. 2006; Characterization of the Vibrio cholerae vexAB and vexCD efflux systems. Arch Microbiol186:171–181
    [Google Scholar]
  5. Cabanillas-Beltran H., LLausás-Magaña E., Romero R., Espinoza A., Garcia-Gasca A., Nishibuchi M., Ishibashi M., Gomez-Gil B.. 2006; Outbreak of gastroenteritis caused by the pandemic Vibrio parahaemolyticus O3 : K6 in Mexico. FEMS Microbiol Lett265:76–80
    [Google Scholar]
  6. CDC 2006; Vibrio parahaemolyticus infections associated with consumption of raw shellfish – three states, 2006. MMWR Morb Mortal Wkly Rep55:854–856
    [Google Scholar]
  7. Chen J., Morita Y., Huda M. N., Kuroda T., Mizushima T., Tsuchiya T.. 2002; VmrA, a member of a novel class of Na+-coupled multidrug efflux pumps from Vibrio parahaemolyticus . J Bacteriol184:572–576
    [Google Scholar]
  8. Chen J., Kuroda T., Huda M. N., Mizushima T., Tsuchiya T.. 2003; An RND-type multidrug efflux pump SdeXY from Serratia marcescens . J Antimicrob Chemother52:176–179
    [Google Scholar]
  9. Cherepanov P. P., Wackernagel W.. 1995; Gene disruption in Escherichia coli : TcR and KmR cassettes with the option of Flp-catalyzed excision of the antibiotic-resistance determinant. Gene158:9–14
    [Google Scholar]
  10. Datsenko K. A., Wanner B. L.. 2000; One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. Proc Natl Acad Sci U S A97:6640–6645
    [Google Scholar]
  11. Fralick J. A.. 1996; Evidence that TolC is required for functioning of the Mar/AcrAB efflux pump of Escherichia coli . J Bacteriol178:5803–5805
    [Google Scholar]
  12. Hamashima H., Iwasaki M., Arai T.. 1995; A simple and rapid method for transformation of Vibrio species by electroporation. Methods Mol Biol47:155–160
    [Google Scholar]
  13. Hanahan D., Jessee J., Bloom F. R.. 1991; Plasmid transformation of Escherichia coli and other bacteria. Methods Enzymol204:63–113
    [Google Scholar]
  14. Hansen L. H., Johannesen E., Burmolle M., Sorensen A. H., Sorensen S. J.. 2004; Plasmid-encoded multidrug efflux pump conferring resistance to olaquindox in Escherichia coli . Antimicrob Agents Chemother48:3332–3337
    [Google Scholar]
  15. Heidelberg J. F., Eisen J. A., Nelson W. C., Clayton R. A., Gwinn M. L., Dodson R. J., Haft D. H., Hickey E. K., Peterson J. D.. other authors 2000; DNA sequence of both chromosomes of the cholera pathogen Vibrio cholerae . Nature406:477–483
    [Google Scholar]
  16. Heidelberg J. F., Paulsen I. T., Nelson K. E., Gaidos E. J., Nelson W. C., Read T. D., Eisen J. A., Seshadri R., Ward N.. other authors 2002; Genome sequence of the dissimilatory metal ion-reducing bacterium Shewanella oneidensis . Nat Biotechnol20:1118–1123
    [Google Scholar]
  17. Henikoff S.. 1984; Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene28:351–359
    [Google Scholar]
  18. Herz K., Vimont S., Padan E., Berche P.. 2003; Roles of NhaA, NhaB, and NhaD Na+/H+ antiporters in survival of Vibrio cholerae in a saline environment. J Bacteriol185:1236–1244
    [Google Scholar]
  19. Huda M. N., Chen J., Morita Y., Kuroda T., Mizushima T., Tsuchiya T.. 2003; Gene cloning and characterization of VcrM, a Na+-coupled multidrug efflux pump, from Vibrio cholerae non-O1. Microbiol Immunol47:419–427
    [Google Scholar]
  20. Japanese Society of Chemotherapy 1990; Microbroth dilution methods for determination of minimum inhibitory concentrations. Chemotherapy38:102–105
    [Google Scholar]
  21. Klein J. R., Henrich B., Plapp R.. 1991; Molecular analysis and nucleotide sequence of the envCD operon of Escherichia coli . Mol Gen Genet230:230–240
    [Google Scholar]
  22. Kumar A., Worobec E. A.. 2005a; Cloning, sequencing, and characterization of the SdeAB multidrug efflux pump of Serratia marcescens . Antimicrob Agents Chemother49:1495–1501
    [Google Scholar]
  23. Kumar A., Worobec E. A.. 2005b; HasF, a TolC-homolog of Serratia marcescens , is involved in energy-dependent efflux. Can J Microbiol51:497–500
    [Google Scholar]
  24. Kumazawa N. H., Kato E.. 1985; Survival of Kanagawa-positive strains of Vibrio parahaemolyticus in a brackish-water area. J Hyg (Lond95:299–307
    [Google Scholar]
  25. Kumazawa N. H., Fukuma N., Komoda Y.. 1991; Attachment of Vibrio parahaemolyticus strains to estuarine algae. J Vet Med Sci53:201–205
    [Google Scholar]
  26. Kuroda T., Mizushima T., Tsuchiya T.. 2005; Physiological roles of three Na+/H+ antiporters in the halophilic bacterium Vibrio parahaemolyticus . Microbiol Immunol49:711–719
    [Google Scholar]
  27. Li X. Z., Nikaido H., Poole K.. 1995; Role of mexA-mexB-oprM in antibiotic efflux in Pseudomonas aeruginosa . Antimicrob Agents Chemother39:1948–1953
    [Google Scholar]
  28. Lozano-Leon A., Torres J., Osorio C. R., Martinez-Urtaza J.. 2003; Identification of tdh-positive Vibrio parahaemolyticus from an outbreak associated with raw oyster consumption in Spain. FEMS Microbiol Lett226:281–284
    [Google Scholar]
  29. Ma D., Cook D. N., Alberti M., Pon N. G., Nikaido H., Hearst J. E.. 1993; Molecular cloning and characterization of acrA and acrE genes of Escherichia coli . J Bacteriol175:6299–6313
    [Google Scholar]
  30. Ma D., Cook D. N., Hearst J. E., Nikaido H.. 1994; Efflux pumps and drug resistance in gram-negative bacteria. Trends Microbiol2:489–493
    [Google Scholar]
  31. Makino K., Oshima K., Kurokawa K., Yokoyama K., Uda T., Tagomori K., Iijima Y., Najima M., Nakano M.. other authors 2003; Genome sequence of Vibrio parahaemolyticus : a pathogenic mechanism distinct from that of V. cholerae . Lancet361:743–749
    [Google Scholar]
  32. Masaoka Y., Ueno Y., Morita Y., Kuroda T., Mizushima T., Tsuchiya T.. 2000; A two-component multidrug efflux pump, EbrAB, in Bacillus subtilis . J Bacteriol182:2307–2310
    [Google Scholar]
  33. McLaughlin J. B., DePaola A., Bopp C. A., Martinek K. A., Napolilli N. P., Allison C. G., Murray S. L., Thompson E. C., Bird M. M., Middaugh J. P.. 2005; Outbreak of Vibrio parahaemolyticus gastroenteritis associated with Alaskan oysters. N Engl J Med353:1463–1470
    [Google Scholar]
  34. Mine T., Morita Y., Kataoka A., Mizushima T., Tsuchiya T.. 1999; Expression in Escherichia coli of a new multidrug efflux pump, MexXY, from Pseudomonas aeruginosa . Antimicrob Agents Chemother43:415–417
    [Google Scholar]
  35. Morita Y., Kodama K., Shiota S., Mine T., Kataoka A., Mizushima T., Tsuchiya T.. 1998; NorM, a putative multidrug efflux protein, of Vibrio parahaemolyticus and its homolog in Escherichia coli . Antimicrob Agents Chemother42:1778–1782
    [Google Scholar]
  36. Morita Y., Kataoka A., Shiota S., Mizushima T., Tsuchiya T.. 2000; NorM of Vibrio parahaemolyticus is an Na+-driven multidrug efflux pump. J Bacteriol182:6694–6697
    [Google Scholar]
  37. Morita Y., Komori Y., Mima T., Kuroda T., Mizushima T., Tsuchiya T.. 2001; Construction of a series of mutants lacking all of the four major mex operons for multidrug efflux pumps or possessing each one of the operons from Pseudomonas aeruginosa PAO1: MexCD-OprJ is an inducible pump. FEMS Microbiol Lett202:139–143
    [Google Scholar]
  38. Murakami S., Nakashima R., Yamashita E., Yamaguchi A.. 2002; Crystal structure of bacterial multidrug efflux transporter AcrB. Nature419:587–593
    [Google Scholar]
  39. Nagakubo S., Nishino K., Hirata T., Yamaguchi A.. 2002; The putative response regulator BaeR stimulates multidrug resistance of Escherichia coli via a novel multidrug exporter system. MdtABC. J Bacteriol184:4161–4167
    [Google Scholar]
  40. Nishino K., Yamaguchi A.. 2001; Analysis of a complete library of putative drug transporter genes in Escherichia coli . J Bacteriol183:5803–5812
    [Google Scholar]
  41. Nishino K., Yamada J., Hirakawa H., Hirata T., Yamaguchi A.. 2003; Roles of TolC-dependent multidrug transporters of Escherichia coli in resistance to β -lactams. Antimicrob Agents Chemother47:3030–3033
    [Google Scholar]
  42. Obata H., Kai A., Morozumi S.. 2001; The trends of Vibrio parahaemolyticus foodborne outbreaks in Tokyo: 1989–2000. Kansenshogaku Zasshi75:485–489
    [Google Scholar]
  43. Ocaktan A., Yoneyama H., Nakae T.. 1997; Use of fluorescence probes to monitor function of the subunit proteins of the MexA-MexB-OprM drug extrusion machinery in Pseudomonas aeruginosa . J Biol Chem272:21964–21969
    [Google Scholar]
  44. Okusu H., Ma D., Nikaido H.. 1996; AcrAB efflux pump plays a major role in the antibiotic resistance phenotype of Escherichia coli multiple-antibiotic-resistance (Mar) mutants. J Bacteriol178:306–308
    [Google Scholar]
  45. Piddock L. J.. 2006; Multidrug-resistance efflux pumps – not just for resistance. Nat Rev Microbiol4:629–636
    [Google Scholar]
  46. Poole K., Krebes K., McNally C., Neshat S.. 1993; Multiple antibiotic resistance in Pseudomonas aeruginosa : evidence for involvement of an efflux operon. J Bacteriol175:7363–7372
    [Google Scholar]
  47. Provenzano D., Schuhmacher D. A., Barker J. L., Klose K. E.. 2000; The virulence regulatory protein ToxR mediates enhanced bile resistance in Vibrio cholerae and other pathogenic Vibrio species. Infect Immun68:1491–1497
    [Google Scholar]
  48. Putman M., van Veen H. W., Konings W. N.. 2000; Molecular properties of bacterial multidrug transporters. Microbiol Mol Biol Rev64:672–693
    [Google Scholar]
  49. Ruby E. G., Urbanowski M., Campbell J., Dunn A., Faini M., Gunsalus R., Lostroh P., Lupp C., McCann J.. other authors 2005; Complete genome sequence of Vibrio fischeri : a symbiotic bacterium with pathogenic congeners. Proc Natl Acad Sci U S A102:3004–3009
    [Google Scholar]
  50. Sekiya H., Mima T., Morita Y., Kuroda T., Mizushima T., Tsuchiya T.. 2003; Functional cloning and characterization of a multidrug efflux pump, mexHI-opmD , from a Pseudomonas aeruginosa mutant. Antimicrob Agents Chemother47:2990–2992
    [Google Scholar]
  51. Sen B., Dutta B., Chatterjee S., Bhattacharya M. K., Nandy R. K., Mukhopadhyay A. K., Gangopadhyay D. N., Bhattacharya S. K., Ramamurthy T.. 2007; The first outbreak of acute diarrhea due to a pandemic strain of Vibrio parahaemolyticus O3 : K6 in Kolkata, India. Int J Infect Dis11:185–187
    [Google Scholar]
  52. Srikumar R., Kon T., Gotoh N., Poole K.. 1998; Expression of Pseudomonas aeruginosa multidrug efflux pumps MexA-MexB-OprM and MexC-MexD-OprJ in a multidrug-sensitive Escherichia coli strain. Antimicrob Agents Chemother42:65–71
    [Google Scholar]
  53. Su H. P., Chiu S. I., Tsai J. L., Lee C. L., Pan T. M.. 2005a; Bacterial food-borne illness outbreaks in northern Taiwan, 1995–2001. J Infect Chemother11:146–151
    [Google Scholar]
  54. Su X. Z., Chen J., Mizushima T., Kuroda T., Tsuchiya T.. 2005b; AbeM, an H+-coupled Acinetobacter baumannii multidrug efflux pump belonging to the MATE family of transporters. Antimicrob Agents Chemother49:4362–4364
    [Google Scholar]
  55. Tamura N., Murakami S., Oyama Y., Ishiguro M., Yamaguchi A.. 2005; Direct interaction of multidrug efflux transporter AcrB and outer membrane channel TolC detected via site-directed disulfide cross-linking. Biochemistry44:11115–11121
    [Google Scholar]
  56. Thanassi D. G., Cheng L. W., Nikaido H.. 1997; Active efflux of bile salts by Escherichia coli . J Bacteriol179:2512–2518
    [Google Scholar]
  57. Tseng T. T., Gratwick K. S., Kollman J., Park D., Nies D. H., Goffeau A., Saier M. H. Jr. 1999; The RND permease superfamily: an ancient, ubiquitous and diverse family that includes human disease and development proteins. J Mol Microbiol Biotechnol1:107–125
    [Google Scholar]
  58. WHO 1999; Vibrio parahaemolyticus , Japan, 1996–1998. Wkly Epidemiol Rec74:361–363
    [Google Scholar]
  59. Xu X. J., Su X. Z., Morita Y., Kuroda T., Mizushima T., Tsuchiya T.. 2003; Molecular cloning and characterization of the HmrM multidrug efflux pump from Haemophilus influenzae Rd. Microbiol Immunol47:937–943
    [Google Scholar]
  60. Yamamoto T., Yokota T.. 1989; Adherence targets of Vibrio parahaemolyticus in human small intestines. Infect Immun57:2410–2419
    [Google Scholar]
  61. Yamazaki M., Inuzuka K., Matsumoto M., Miwa Y., Hiramatsu R., Matsui H., Sakae K., Suzuki Y., Miyazaki Y.. 2003; Epidemiological study of outbreaks and sporadic cases due to Vibrio parahaemolyticus – serotype O3 : K6 in Aichi Prefecture, Japan, during 1988 and 2001. Kansenshogaku Zasshi77:1015–1023
    [Google Scholar]
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