Skip to main content
Antimicrobial Agents and Chemotherapy logoLink to Antimicrobial Agents and Chemotherapy
. 1995 Feb;39(2):453–457. doi: 10.1128/aac.39.2.453

Cloning of multidrug resistance gene pqrA from Proteus vulgaris.

H Ishida 1, H Fuziwara 1, Y Kaibori 1, T Horiuchi 1, K Sato 1, Y Osada 1
PMCID: PMC162559  PMID: 7726514

Abstract

The multiple antibiotic resistance gene pqrA was cloned from the chromosomal DNA of a clinical isolate of Proteus vulgaris 881051 into Escherichia coli KY2563. The MICs of quinolones tetracycline, cephalosporin, and chloramphenicol for transformant strain DNS7020 were from 8 to 32 times higher than those for the parent strain, KY2563. The level of expression of outer membrane protein F (OmpF) by DNS7020 was lower than that of KY2563 but not as low as that of an OmpF-deficient control strain. The 1.4-kb fragment containing the pqrA gene had an open reading frame encoding a polypeptide of 122 amino acid residues with a molecular weight of about 14,000, which was consistent with the experimental value identified by the Maxicell method. The putative PqrA polypeptide showed significant amino acid sequence similarity to the E. coli proteins SoxS and MarA. These polypeptides are strongly conserved in predicted helix-turn-helix DNA binding domains. The MarA protein, which is responsible for multiple antibiotic resistance in E. coli, also decreases OmpF expression. Moreover, the SoxS protein, which is characterized as a superoxide response regulon of E. coli, has also been shown to increase resistance to many structurally unrelated antibiotics. The soxS gene increases superoxide dismutase levels in addition to decreasing OmpF expression. The expression level of superoxide dismutase with DNS7020 was about 1.5 times higher than that with KY2563. These findings suggest that the pqrA gene in P. vulgaris confers multidrug resistance in a way similar to that of the soxS and marA genes in E. coli.

Full Text

The Full Text of this article is available as a PDF (242.9 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Acar J. F., Francoual S. The clinical problems of bacterial resistance to the new quinolones. J Antimicrob Chemother. 1990 Oct;26 (Suppl B):207–213. doi: 10.1093/jac/26.suppl_b.207. [DOI] [PubMed] [Google Scholar]
  2. Amábile-Cuevas C. F., Demple B. Molecular characterization of the soxRS genes of Escherichia coli: two genes control a superoxide stress regulon. Nucleic Acids Res. 1991 Aug 25;19(16):4479–4484. doi: 10.1093/nar/19.16.4479. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Appelbaum P. C. Mechanisms and frequency of resistance to temafloxacin. Am J Med. 1991 Dec 30;91(6A):27S–30S. doi: 10.1016/0002-9343(91)90306-i. [DOI] [PubMed] [Google Scholar]
  4. Ariza R. R., Cohen S. P., Bachhawat N., Levy S. B., Demple B. Repressor mutations in the marRAB operon that activate oxidative stress genes and multiple antibiotic resistance in Escherichia coli. J Bacteriol. 1994 Jan;176(1):143–148. doi: 10.1128/jb.176.1.143-148.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bryan L. E., Bedard J. Impermeability to quinolones in gram-positive and gram-negative bacteria. Eur J Clin Microbiol Infect Dis. 1991 Apr;10(4):232–239. doi: 10.1007/BF01966995. [DOI] [PubMed] [Google Scholar]
  6. Bryan L. E., Bedard J., Wong S., Chamberland S. Quinolone antimicrobial agents: mechanism of action and resistance development. Clin Invest Med. 1989 Feb;12(1):14–19. [PubMed] [Google Scholar]
  7. Cohen S. P., Hächler H., Levy S. B. Genetic and functional analysis of the multiple antibiotic resistance (mar) locus in Escherichia coli. J Bacteriol. 1993 Mar;175(5):1484–1492. doi: 10.1128/jb.175.5.1484-1492.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cohen S. P., McMurry L. M., Hooper D. C., Wolfson J. S., Levy S. B. Cross-resistance to fluoroquinolones in multiple-antibiotic-resistant (Mar) Escherichia coli selected by tetracycline or chloramphenicol: decreased drug accumulation associated with membrane changes in addition to OmpF reduction. Antimicrob Agents Chemother. 1989 Aug;33(8):1318–1325. doi: 10.1128/aac.33.8.1318. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Cohen S. P., McMurry L. M., Levy S. B. marA locus causes decreased expression of OmpF porin in multiple-antibiotic-resistant (Mar) mutants of Escherichia coli. J Bacteriol. 1988 Dec;170(12):5416–5422. doi: 10.1128/jb.170.12.5416-5422.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Dang P., Gutmann L., Quentin C., Williamson R., Collatz E. Some properties of Serratia marcescens, Salmonella paratyphi A, and Enterobacter cloacae with non-enzyme-dependent multiple resistance to beta-lactam antibiotics, aminoglycosides, and quinolones. Rev Infect Dis. 1988 Jul-Aug;10(4):899–904. doi: 10.1093/clinids/10.4.899. [DOI] [PubMed] [Google Scholar]
  11. Dechène M., Leying H., Cullmann W. Role of the outer membrane for quinolone resistance in enterobacteria. Chemotherapy. 1990;36(1):13–23. doi: 10.1159/000238743. [DOI] [PubMed] [Google Scholar]
  12. Del Sal G., Manfioletti G., Schneider C. A one-tube plasmid DNA mini-preparation suitable for sequencing. Nucleic Acids Res. 1988 Oct 25;16(20):9878–9878. doi: 10.1093/nar/16.20.9878. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gambino L., Gracheck S. J., Miller P. F. Overexpression of the MarA positive regulator is sufficient to confer multiple antibiotic resistance in Escherichia coli. J Bacteriol. 1993 May;175(10):2888–2894. doi: 10.1128/jb.175.10.2888-2894.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Greenberg J. T., Chou J. H., Monach P. A., Demple B. Activation of oxidative stress genes by mutations at the soxQ/cfxB/marA locus of Escherichia coli. J Bacteriol. 1991 Jul;173(14):4433–4439. doi: 10.1128/jb.173.14.4433-4439.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hirai K., Aoyama H., Suzue S., Irikura T., Iyobe S., Mitsuhashi S. Isolation and characterization of norfloxacin-resistant mutants of Escherichia coli K-12. Antimicrob Agents Chemother. 1986 Aug;30(2):248–253. doi: 10.1128/aac.30.2.248. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hooper D. C., Wolfson J. S. Bacterial resistance to the quinolone antimicrobial agents. Am J Med. 1989 Dec 29;87(6C):17S–23S. [PubMed] [Google Scholar]
  17. Hooper D. C., Wolfson J. S., Bozza M. A., Ng E. Y. Genetics and regulation of outer membrane protein expression by quinolone resistance loci nfxB, nfxC, and cfxB. Antimicrob Agents Chemother. 1992 May;36(5):1151–1154. doi: 10.1128/aac.36.5.1151. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hooper D. C., Wolfson J. S., Ng E. Y., Swartz M. N. Mechanisms of action of and resistance to ciprofloxacin. Am J Med. 1987 Apr 27;82(4A):12–20. [PubMed] [Google Scholar]
  19. Hooper D. C., Wolfson J. S., Souza K. S., Ng E. Y., McHugh G. L., Swartz M. N. Mechanisms of quinolone resistance in Escherichia coli: characterization of nfxB and cfxB, two mutant resistance loci decreasing norfloxacin accumulation. Antimicrob Agents Chemother. 1989 Mar;33(3):283–290. doi: 10.1128/aac.33.3.283. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hooper D. C., Wolfson J. S., Souza K. S., Tung C., McHugh G. L., Swartz M. N. Genetic and biochemical characterization of norfloxacin resistance in Escherichia coli. Antimicrob Agents Chemother. 1986 Apr;29(4):639–644. doi: 10.1128/aac.29.4.639. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Ishii H., Sato K., Hoshino K., Sato M., Yamaguchi A., Sawai T., Osada Y. Active efflux of ofloxacin by a highly quinolone-resistant strain of Proteus vulgaris. J Antimicrob Chemother. 1991 Dec;28(6):827–836. doi: 10.1093/jac/28.6.827. [DOI] [PubMed] [Google Scholar]
  22. Lewin C. S., Allen R. A., Amyes S. G. Potential mechanisms of resistance to the modern fluorinated 4-quinolones. J Med Microbiol. 1990 Mar;31(3):153–162. doi: 10.1099/00222615-31-3-153. [DOI] [PubMed] [Google Scholar]
  23. Matsuhashi M., Song M. D., Ishino F., Wachi M., Doi M., Inoue M., Ubukata K., Yamashita N., Konno M. Molecular cloning of the gene of a penicillin-binding protein supposed to cause high resistance to beta-lactam antibiotics in Staphylococcus aureus. J Bacteriol. 1986 Sep;167(3):975–980. doi: 10.1128/jb.167.3.975-980.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. McCord J. M., Fridovich I. Superoxide dismutase. An enzymic function for erythrocuprein (hemocuprein). J Biol Chem. 1969 Nov 25;244(22):6049–6055. [PubMed] [Google Scholar]
  25. Mizushima S., Yamada H. Isolation and characterization of two outer membrane preparations from Escherichia coli. Biochim Biophys Acta. 1975 Jan 14;375(1):44–53. doi: 10.1016/0005-2736(75)90071-1. [DOI] [PubMed] [Google Scholar]
  26. Neu H. C. Quinolone antimicrobial agents. Annu Rev Med. 1992;43:465–486. doi: 10.1146/annurev.me.43.020192.002341. [DOI] [PubMed] [Google Scholar]
  27. Neyfakh A. A., Borsch C. M., Kaatz G. W. Fluoroquinolone resistance protein NorA of Staphylococcus aureus is a multidrug efflux transporter. Antimicrob Agents Chemother. 1993 Jan;37(1):128–129. doi: 10.1128/aac.37.1.128. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Neyfakh A. A. The multidrug efflux transporter of Bacillus subtilis is a structural and functional homolog of the Staphylococcus NorA protein. Antimicrob Agents Chemother. 1992 Feb;36(2):484–485. doi: 10.1128/aac.36.2.484. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Oram M., Fisher L. M. 4-Quinolone resistance mutations in the DNA gyrase of Escherichia coli clinical isolates identified by using the polymerase chain reaction. Antimicrob Agents Chemother. 1991 Feb;35(2):387–389. doi: 10.1128/aac.35.2.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Power E. G., Muñoz Bellido J. L., Phillips I. Detection of ciprofloxacin resistance in gram-negative bacteria due to alterations in gyrA. J Antimicrob Chemother. 1992 Jan;29(1):9–17. doi: 10.1093/jac/29.1.9. [DOI] [PubMed] [Google Scholar]
  31. Roberts C. M., Batten J., Hodson M. E. Ciprofloxacin-resistant Pseudomonas. Lancet. 1985 Jun 22;1(8443):1442–1442. doi: 10.1016/s0140-6736(85)91862-8. [DOI] [PubMed] [Google Scholar]
  32. Sancar A., Hack A. M., Rupp W. D. Simple method for identification of plasmid-coded proteins. J Bacteriol. 1979 Jan;137(1):692–693. doi: 10.1128/jb.137.1.692-693.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Sawai T., Hiruma R., Kawana N., Kaneko M., Taniyasu F., Inami A. Outer membrane permeation of beta-lactam antibiotics in Escherichia coli, Proteus mirabilis, and Enterobacter cloacae. Antimicrob Agents Chemother. 1982 Oct;22(4):585–592. doi: 10.1128/aac.22.4.585. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Trucksis M., Wolfson J. S., Hooper D. C. A novel locus conferring fluoroquinolone resistance in Staphylococcus aureus. J Bacteriol. 1991 Sep;173(18):5854–5860. doi: 10.1128/jb.173.18.5854-5860.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Ubukata K., Itoh-Yamashita N., Konno M. Cloning and expression of the norA gene for fluoroquinolone resistance in Staphylococcus aureus. Antimicrob Agents Chemother. 1989 Sep;33(9):1535–1539. doi: 10.1128/aac.33.9.1535. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Wu J., Weiss B. Two divergently transcribed genes, soxR and soxS, control a superoxide response regulon of Escherichia coli. J Bacteriol. 1991 May;173(9):2864–2871. doi: 10.1128/jb.173.9.2864-2871.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Yoshida H., Bogaki M., Nakamura S., Ubukata K., Konno M. Nucleotide sequence and characterization of the Staphylococcus aureus norA gene, which confers resistance to quinolones. J Bacteriol. 1990 Dec;172(12):6942–6949. doi: 10.1128/jb.172.12.6942-6949.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Antimicrobial Agents and Chemotherapy are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES