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. 1995 Sep;39(9):2145–2149. doi: 10.1128/aac.39.9.2145

Sequences of conserved region in the A subunit of DNA gyrase from nine species of the genus Mycobacterium: phylogenetic analysis and implication for intrinsic susceptibility to quinolones.

I Guillemin 1, E Cambau 1, V Jarlier 1
PMCID: PMC162899  PMID: 8540734

Abstract

The sequences of a conserved region in the A subunit of DNA gyrase corresponding to the quinolone resistance-determining region were determined for nine mycobacterial species and were compared. Although the nucleotide sequences were highly conserved, they clearly differentiated one species from another. The results of the phylogenetic analysis based on the sequences of the quinolone resistance-determining regions were compared with those provided by the 16S rRNA sequences. Deduced amino acid sequences were identical within the nine species except for amino acid 83, which was frequently involved in acquired resistance to quinolones in many genera, including mycobacteria. The presence at position 83 of an alanine for seven mycobacterial species (M. tuberculosis, M. bovis BCG, M. leprae, M. avium, M. kansasii, M. chelonae, and M. smegmatis) and of a serine for the two remaining mycobacterial species (M. fortuitum and M. aurum) correlated well with the MICs of ofloxacin for both groups of species, suggesting the role of this residue in intrinsic susceptibility to quinolones in mycobacteria.

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Selected References

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  1. Austin C. A., Sng J. H., Patel S., Fisher L. M. Novel HeLa topoisomerase II is the II beta isoform: complete coding sequence and homology with other type II topoisomerases. Biochim Biophys Acta. 1993 Mar 20;1172(3):283–291. doi: 10.1016/0167-4781(93)90215-y. [DOI] [PubMed] [Google Scholar]
  2. Cambau E., Gutmann L. Mechanisms of resistance to quinolones. Drugs. 1993;45 (Suppl 3):15–23. doi: 10.2165/00003495-199300453-00005. [DOI] [PubMed] [Google Scholar]
  3. Cambau E., Sougakoff W., Besson M., Truffot-Pernot C., Grosset J., Jarlier V. Selection of a gyrA mutant of Mycobacterium tuberculosis resistant to fluoroquinolones during treatment with ofloxacin. J Infect Dis. 1994 Aug;170(2):479–483. doi: 10.1093/infdis/170.2.479. [DOI] [PubMed] [Google Scholar]
  4. Cambau E., Sougakoff W., Jarlier V. Amplification and nucleotide sequence of the quinolone resistance-determining region in the gyrA gene of mycobacteria. FEMS Microbiol Lett. 1994 Feb 1;116(1):49–54. doi: 10.1111/j.1574-6968.1994.tb06674.x. [DOI] [PubMed] [Google Scholar]
  5. Cullen M. E., Wyke A. W., Kuroda R., Fisher L. M. Cloning and characterization of a DNA gyrase A gene from Escherichia coli that confers clinical resistance to 4-quinolones. Antimicrob Agents Chemother. 1989 Jun;33(6):886–894. doi: 10.1128/aac.33.6.886. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gellert M., Mizuuchi K., O'Dea M. H., Itoh T., Tomizawa J. I. Nalidixic acid resistance: a second genetic character involved in DNA gyrase activity. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4772–4776. doi: 10.1073/pnas.74.11.4772. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gellert M., Mizuuchi K., O'Dea M. H., Nash H. A. DNA gyrase: an enzyme that introduces superhelical turns into DNA. Proc Natl Acad Sci U S A. 1976 Nov;73(11):3872–3876. doi: 10.1073/pnas.73.11.3872. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hallett P., Maxwell A. Novel quinolone resistance mutations of the Escherichia coli DNA gyrase A protein: enzymatic analysis of the mutant proteins. Antimicrob Agents Chemother. 1991 Feb;35(2):335–340. doi: 10.1128/aac.35.2.335. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hopewell R., Oram M., Briesewitz R., Fisher L. M. DNA cloning and organization of the Staphylococcus aureus gyrA and gyrB genes: close homology among gyrase proteins and implications for 4-quinolone action and resistance. J Bacteriol. 1990 Jun;172(6):3481–3484. doi: 10.1128/jb.172.6.3481-3484.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Horowitz D. S., Wang J. C. Mapping the active site tyrosine of Escherichia coli DNA gyrase. J Biol Chem. 1987 Apr 15;262(11):5339–5344. [PubMed] [Google Scholar]
  11. Jarlier V., Nikaido H. Mycobacterial cell wall: structure and role in natural resistance to antibiotics. FEMS Microbiol Lett. 1994 Oct 15;123(1-2):11–18. doi: 10.1111/j.1574-6968.1994.tb07194.x. [DOI] [PubMed] [Google Scholar]
  12. Jarlier V., Nikaido H. Permeability barrier to hydrophilic solutes in Mycobacterium chelonei. J Bacteriol. 1990 Mar;172(3):1418–1423. doi: 10.1128/jb.172.3.1418-1423.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Ji B., Perani E. G., Petinon C., Grosset J. H. Bactericidal activities of single or multiple doses of various combinations of new antileprosy drugs and/or rifampin against M. leprae in mice. Int J Lepr Other Mycobact Dis. 1992 Dec;60(4):556–561. [PubMed] [Google Scholar]
  14. Kureishi A., Diver J. M., Beckthold B., Schollaardt T., Bryan L. E. Cloning and nucleotide sequence of Pseudomonas aeruginosa DNA gyrase gyrA gene from strain PAO1 and quinolone-resistant clinical isolates. Antimicrob Agents Chemother. 1994 Sep;38(9):1944–1952. doi: 10.1128/aac.38.9.1944. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Leysen D. C., Haemers A., Pattyn S. R. Mycobacteria and the new quinolones. Antimicrob Agents Chemother. 1989 Jan;33(1):1–5. doi: 10.1128/aac.33.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lounis N., Ji B., Truffot-Pernot C., Grosset J. Selection of clarithromycin-resistant Mycobacterium avium complex during combined therapy using the beige mouse model. Antimicrob Agents Chemother. 1995 Mar;39(3):608–612. doi: 10.1128/AAC.39.3.608. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Margerrison E. E., Hopewell R., Fisher L. M. Nucleotide sequence of the Staphylococcus aureus gyrB-gyrA locus encoding the DNA gyrase A and B proteins. J Bacteriol. 1992 Mar;174(5):1596–1603. doi: 10.1128/jb.174.5.1596-1603.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Moriya S., Ogasawara N., Yoshikawa H. Structure and function of the region of the replication origin of the Bacillus subtilis chromosome. III. Nucleotide sequence of some 10,000 base pairs in the origin region. Nucleic Acids Res. 1985 Apr 11;13(7):2251–2265. doi: 10.1093/nar/13.7.2251. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Revel V., Cambau E., Jarlier V., Sougakoff W. Characterization of mutations in Mycobacterium smegmatis involved in resistance to fluoroquinolones. Antimicrob Agents Chemother. 1994 Sep;38(9):1991–1996. doi: 10.1128/aac.38.9.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Rogall T., Flohr T., Böttger E. C. Differentiation of Mycobacterium species by direct sequencing of amplified DNA. J Gen Microbiol. 1990 Sep;136(9):1915–1920. doi: 10.1099/00221287-136-9-1915. [DOI] [PubMed] [Google Scholar]
  21. Saitou N., Nei M. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol. 1987 Jul;4(4):406–425. doi: 10.1093/oxfordjournals.molbev.a040454. [DOI] [PubMed] [Google Scholar]
  22. Snapper S. B., Melton R. E., Mustafa S., Kieser T., Jacobs W. R., Jr Isolation and characterization of efficient plasmid transformation mutants of Mycobacterium smegmatis. Mol Microbiol. 1990 Nov;4(11):1911–1919. doi: 10.1111/j.1365-2958.1990.tb02040.x. [DOI] [PubMed] [Google Scholar]
  23. Sreedharan S., Oram M., Jensen B., Peterson L. R., Fisher L. M. DNA gyrase gyrA mutations in ciprofloxacin-resistant strains of Staphylococcus aureus: close similarity with quinolone resistance mutations in Escherichia coli. J Bacteriol. 1990 Dec;172(12):7260–7262. doi: 10.1128/jb.172.12.7260-7262.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Stackebrandt E., Smida J., Kazda J. The primary structure of the 16SrRNA of Mycobacterium leprae: its use in phylogeny and development of DNA probes. Acta Leprol. 1989;7 (Suppl 1):222–225. [PubMed] [Google Scholar]
  25. Stahl D. A., Urbance J. W. The division between fast- and slow-growing species corresponds to natural relationships among the mycobacteria. J Bacteriol. 1990 Jan;172(1):116–124. doi: 10.1128/jb.172.1.116-124.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Steele L. C., Wallace R. J., Jr Ability of ciprofloxacin but not pipemidic acid to differentiate all three biovariants of Mycobacterium fortuitum from Mycobacterium chelonae. J Clin Microbiol. 1987 Feb;25(2):456–457. doi: 10.1128/jcm.25.2.456-457.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Takiff H. E., Salazar L., Guerrero C., Philipp W., Huang W. M., Kreiswirth B., Cole S. T., Jacobs W. R., Jr, Telenti A. Cloning and nucleotide sequence of Mycobacterium tuberculosis gyrA and gyrB genes and detection of quinolone resistance mutations. Antimicrob Agents Chemother. 1994 Apr;38(4):773–780. doi: 10.1128/aac.38.4.773. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Tsukamura M., Nakamura E., Yoshii S., Amano H. Therapeutic effect of a new antibacterial substance ofloxacin (DL8280) on pulmonary tuberculosis. Am Rev Respir Dis. 1985 Mar;131(3):352–356. doi: 10.1164/arrd.1985.131.3.352. [DOI] [PubMed] [Google Scholar]
  29. Wallace R. J., Jr, Bedsole G., Sumter G., Sanders C. V., Steele L. C., Brown B. A., Smith J., Graham D. R. Activities of ciprofloxacin and ofloxacin against rapidly growing mycobacteria with demonstration of acquired resistance following single-drug therapy. Antimicrob Agents Chemother. 1990 Jan;34(1):65–70. doi: 10.1128/aac.34.1.65. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Wang Y., Huang W. M., Taylor D. E. Cloning and nucleotide sequence of the Campylobacter jejuni gyrA gene and characterization of quinolone resistance mutations. Antimicrob Agents Chemother. 1993 Mar;37(3):457–463. doi: 10.1128/aac.37.3.457. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Willmott C. J., Maxwell A. A single point mutation in the DNA gyrase A protein greatly reduces binding of fluoroquinolones to the gyrase-DNA complex. Antimicrob Agents Chemother. 1993 Jan;37(1):126–127. doi: 10.1128/aac.37.1.126. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Woods S. A., Cole S. T. A rapid method for the detection of potentially viable Mycobacterium leprae in human biopsies: a novel application of PCR. FEMS Microbiol Lett. 1989 Dec;53(3):305–309. doi: 10.1016/0378-1097(89)90235-8. [DOI] [PubMed] [Google Scholar]
  33. Yoshida H., Bogaki M., Nakamura M., Nakamura S. Quinolone resistance-determining region in the DNA gyrase gyrA gene of Escherichia coli. Antimicrob Agents Chemother. 1990 Jun;34(6):1271–1272. doi: 10.1128/aac.34.6.1271. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. van Soolingen D., Hermans P. W., de Haas P. E., Soll D. R., van Embden J. D. Occurrence and stability of insertion sequences in Mycobacterium tuberculosis complex strains: evaluation of an insertion sequence-dependent DNA polymorphism as a tool in the epidemiology of tuberculosis. J Clin Microbiol. 1991 Nov;29(11):2578–2586. doi: 10.1128/jcm.29.11.2578-2586.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]

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