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. 1995 Nov;39(11):2392–2396. doi: 10.1128/aac.39.11.2392

OprK and OprM define two genetically distinct multidrug efflux systems in Pseudomonas aeruginosa.

M M Hamzehpour 1, J C Pechere 1, P Plesiat 1, T Köhler 1
PMCID: PMC162953  PMID: 8585714

Abstract

Multidrug-resistant derivatives of Pseudomonas aeruginosa PAO1 were obtained after stepwise selection on tetracycline or erythromycin. Two phenotypes were generated. The tetracycline-resistant mutant (TETR) was phenotypically similar to OprM-overexpressing strains. This group displayed cross-resistance to quinolones, chloramphenicol, and all beta-lactams tested except imipenem, with no changes in the erythromycin MICs for the strains. Sodium dodecyl sulfate-polyacrylamide gels showed the overproduction of an outer membrane protein in the range of 50 kDa and a 46-kDa inner membrane protein. The erythromycin-resistant mutant (ERYR) kept its susceptibility to all beta-lactams tested with the exception of cefpirome, but it was resistant to chloramphenicol, quinolones, and tetracycline and was hypersusceptible to imipenem. This mutant also exhibited overexpression of a 50-kDa outer membrane protein that was different from OprM and of a 43-kDa inner membrane protein. The phenotype of ERYR was comparable to those of OprK- and OprJ-overexpressing strains. These strains were therefore classified as the OprK-like group. Transduction of the oprK::omega-Hg mutation of strain K613 (K. Poole, K. Krebes, C. McNally, and S. Neshat, J. Bacteriol. 175:7363-7372, 1993) into the multidrug-resistant strains resulted in the loss of multidrug resistance and the acquisition of hypersusceptibility in the OprM group, while the phenotype of the OprK-like group was unaffected. These experiments demonstrated the existence of two genetically distinct efflux systems in P. aeruginosa. The identities of the operons encoding the two efflux systems and their physiological roles are discussed.

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Selected References

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  1. Bryson V., Szybalski W. Microbial Selection. Science. 1952 Jul 18;116(3003):45–51. doi: 10.1126/science.116.3003.45. [DOI] [PubMed] [Google Scholar]
  2. Fellay R., Frey J., Krisch H. Interposon mutagenesis of soil and water bacteria: a family of DNA fragments designed for in vitro insertional mutagenesis of gram-negative bacteria. Gene. 1987;52(2-3):147–154. doi: 10.1016/0378-1119(87)90041-2. [DOI] [PubMed] [Google Scholar]
  3. Fukuda H., Hosaka M., Hirai K., Iyobe S. New norfloxacin resistance gene in Pseudomonas aeruginosa PAO. Antimicrob Agents Chemother. 1990 Sep;34(9):1757–1761. doi: 10.1128/aac.34.9.1757. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Gotoh N., Itoh N., Tsujimoto H., Yamagishi J., Oyamada Y., Nishino T. Isolation of OprM-deficient mutants of Pseudomonas aeruginosa by transposon insertion mutagenesis: evidence of involvement in multiple antibiotic resistance. FEMS Microbiol Lett. 1994 Oct 1;122(3):267–273. doi: 10.1111/j.1574-6968.1994.tb07179.x. [DOI] [PubMed] [Google Scholar]
  5. Haas D., Holloway B. W., Schamböck A., Leisinger T. The genetic organization of arginine biosynthesis in Pseudomonas aeruginosa. Mol Gen Genet. 1977 Jul 7;154(1):7–22. doi: 10.1007/BF00265571. [DOI] [PubMed] [Google Scholar]
  6. Hirai K., Suzue S., Irikura T., Iyobe S., Mitsuhashi S. Mutations producing resistance to norfloxacin in Pseudomonas aeruginosa. Antimicrob Agents Chemother. 1987 Apr;31(4):582–586. doi: 10.1128/aac.31.4.582. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Levy S. B. Active efflux mechanisms for antimicrobial resistance. Antimicrob Agents Chemother. 1992 Apr;36(4):695–703. doi: 10.1128/aac.36.4.695. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Lewis K. Multidrug resistance pumps in bacteria: variations on a theme. Trends Biochem Sci. 1994 Mar;19(3):119–123. doi: 10.1016/0968-0004(94)90204-6. [DOI] [PubMed] [Google Scholar]
  9. Li X. Z., Livermore D. M., Nikaido H. Role of efflux pump(s) in intrinsic resistance of Pseudomonas aeruginosa: resistance to tetracycline, chloramphenicol, and norfloxacin. Antimicrob Agents Chemother. 1994 Aug;38(8):1732–1741. doi: 10.1128/aac.38.8.1732. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Li X. Z., Ma D., Livermore D. M., Nikaido H. Role of efflux pump(s) in intrinsic resistance of Pseudomonas aeruginosa: active efflux as a contributing factor to beta-lactam resistance. Antimicrob Agents Chemother. 1994 Aug;38(8):1742–1752. doi: 10.1128/aac.38.8.1742. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Masuda N., Ohya S. Cross-resistance to meropenem, cephems, and quinolones in Pseudomonas aeruginosa. Antimicrob Agents Chemother. 1992 Sep;36(9):1847–1851. doi: 10.1128/aac.36.9.1847. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Masuda N., Sakagawa E., Ohya S. Outer membrane proteins responsible for multiple drug resistance in Pseudomonas aeruginosa. Antimicrob Agents Chemother. 1995 Mar;39(3):645–649. doi: 10.1128/AAC.39.3.645. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Michéa-Hamzehpour M., Pechère J. C., Marchou B., Auckenthaler R. Combination therapy: a way to limit emergence of resistance? Am J Med. 1986 Jun 30;80(6B):138–142. doi: 10.1016/0002-9343(86)90491-2. [DOI] [PubMed] [Google Scholar]
  14. Morgan A. F. Transduction of Pseudomonas aeruginosa with a mutant of bacteriophage E79. J Bacteriol. 1979 Jul;139(1):137–140. doi: 10.1128/jb.139.1.137-140.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Nikaido H. Prevention of drug access to bacterial targets: permeability barriers and active efflux. Science. 1994 Apr 15;264(5157):382–388. doi: 10.1126/science.8153625. [DOI] [PubMed] [Google Scholar]
  16. Poole K., Heinrichs D. E., Neshat S. Cloning and sequence analysis of an EnvCD homologue in Pseudomonas aeruginosa: regulation by iron and possible involvement in the secretion of the siderophore pyoverdine. Mol Microbiol. 1993 Nov;10(3):529–544. doi: 10.1111/j.1365-2958.1993.tb00925.x. [DOI] [PubMed] [Google Scholar]
  17. Poole K., Krebes K., McNally C., Neshat S. Multiple antibiotic resistance in Pseudomonas aeruginosa: evidence for involvement of an efflux operon. J Bacteriol. 1993 Nov;175(22):7363–7372. doi: 10.1128/jb.175.22.7363-7372.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Poole K., Neshat S., Heinrichs D. Pyoverdine-mediated iron transport in Pseudomonas aeruginosa: involvement of a high-molecular-mass outer membrane protein. FEMS Microbiol Lett. 1991 Feb;62(1):1–5. [PubMed] [Google Scholar]
  19. Rella M., Haas D. Resistance of Pseudomonas aeruginosa PAO to nalidixic acid and low levels of beta-lactam antibiotics: mapping of chromosomal genes. Antimicrob Agents Chemother. 1982 Aug;22(2):242–249. doi: 10.1128/aac.22.2.242. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Robillard N. J., Scarpa A. L. Genetic and physiological characterization of ciprofloxacin resistance in Pseudomonas aeruginosa PAO. Antimicrob Agents Chemother. 1988 Apr;32(4):535–539. doi: 10.1128/aac.32.4.535. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]

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