Skip to main content
PMC home page
Antimicrobial Agents and Chemotherapy logoLink to Antimicrobial Agents and Chemotherapy
. 1996 May;40(5):1126–1133. doi: 10.1128/aac.40.5.1126

Cloning and nucleotide sequence of the DNA gyrase gyrA gene from the fish pathogen Aeromonas salmonicida.

H Oppegaard 1, H Sørum 1
PMCID: PMC163277  PMID: 8723452

Abstract

The DNA gyrase gyrA gene from the fish pathogen Aeromonas salmonicida 2148/89 was cloned, and the nucleotide sequence was determined. An open reading frame of 2,766 nucleotides was identified and was found to encode a protein of 922 amino acids with a calculated molecular mass of 101.1 kDa. The derived amino acid sequence shared a high degree of identity with other DNA gyrase A proteins, in particular, with other gram-negative GyrA sequences. When the amino acid sequence of A. salmonicida GyrA was compared with that of Escherichia coli GyrA, a number of conserved residues were present at identical coordinates, including the catalytic Tyr residue at position 122 (Tyr-122) and residues whose substitution confers quinolone resistance, notably, Ser-83, Ala-67, Gly-81, Asp-87, Ala-84, and Gln-106. An intragenic region corresponding to 48 amino acids, which is not present in E. coli or other bacteria, was identified in the C-terminal part of A. salmonicida GyrA. This intragenic region shared sequence identity with various DNA-binding proteins of both prokaryotic and eukaryotic origins.

Full Text

The Full Text of this article is available as a PDF (420.2 KB).

Selected References

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

  1. Ansorge W., Sproat B., Stegemann J., Schwager C., Zenke M. Automated DNA sequencing: ultrasensitive detection of fluorescent bands during electrophoresis. Nucleic Acids Res. 1987 Jun 11;15(11):4593–4602. doi: 10.1093/nar/15.11.4593. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bachmann B. J., Low K. B. Linkage map of Escherichia coli K-12, edition 6. Microbiol Rev. 1980 Mar;44(1):1–56. doi: 10.1128/mr.44.1.1-56.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Belland R. J., Morrison S. G., Ison C., Huang W. M. Neisseria gonorrhoeae acquires mutations in analogous regions of gyrA and parC in fluoroquinolone-resistant isolates. Mol Microbiol. 1994 Oct;14(2):371–380. doi: 10.1111/j.1365-2958.1994.tb01297.x. [DOI] [PubMed] [Google Scholar]
  4. Colman S. D., Hu P. C., Bott K. F. Mycoplasma pneumoniae DNA gyrase genes. Mol Microbiol. 1990 Jul;4(7):1129–1134. doi: 10.1111/j.1365-2958.1990.tb00687.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. Deguchi T., Yasuda M., Asano M., Tada K., Iwata H., Komeda H., Ezaki T., Saito I., Kawada Y. DNA gyrase mutations in quinolone-resistant clinical isolates of Neisseria gonorrhoeae. Antimicrob Agents Chemother. 1995 Feb;39(2):561–563. doi: 10.1128/aac.39.2.561. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dimri G. P., Das H. K. Cloning and sequence analysis of gyrA gene of Klebsiella pneumoniae. Nucleic Acids Res. 1990 Jan 11;18(1):151–156. doi: 10.1093/nar/18.1.151. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Ferrero L., Cameron B., Manse B., Lagneaux D., Crouzet J., Famechon A., Blanche F. Cloning and primary structure of Staphylococcus aureus DNA topoisomerase IV: a primary target of fluoroquinolones. Mol Microbiol. 1994 Aug;13(4):641–653. doi: 10.1111/j.1365-2958.1994.tb00458.x. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Horwitz M. S., Loeb L. A. An E. coli promoter that regulates transcription by DNA superhelix-induced cruciform extrusion. Science. 1988 Aug 5;241(4866):703–705. doi: 10.1126/science.2456617. [DOI] [PubMed] [Google Scholar]
  11. Kato J., Suzuki H., Ikeda H. Purification and characterization of DNA topoisomerase IV in Escherichia coli. J Biol Chem. 1992 Dec 25;267(36):25676–25684. [PubMed] [Google Scholar]
  12. Korten V., Huang W. M., Murray B. E. Analysis by PCR and direct DNA sequencing of gyrA mutations associated with fluoroquinolone resistance in Enterococcus faecalis. Antimicrob Agents Chemother. 1994 Sep;38(9):2091–2094. doi: 10.1128/aac.38.9.2091. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kristensen T., Voss H., Schwager C., Stegemann J., Sproat B., Ansorge W. T7 DNA polymerase in automated dideoxy sequencing. Nucleic Acids Res. 1988 Apr 25;16(8):3487–3496. doi: 10.1093/nar/16.8.3487. [DOI] [PMC free article] [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. 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]
  16. Menzel R., Gellert M. Modulation of transcription by DNA supercoiling: a deletion analysis of the Escherichia coli gyrA and gyrB promoters. Proc Natl Acad Sci U S A. 1987 Jun;84(12):4185–4189. doi: 10.1073/pnas.84.12.4185. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Moore R. A., Beckthold B., Wong S., Kureishi A., Bryan L. E. Nucleotide sequence of the gyrA gene and characterization of ciprofloxacin-resistant mutants of Helicobacter pylori. Antimicrob Agents Chemother. 1995 Jan;39(1):107–111. doi: 10.1128/aac.39.1.107. [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. Oppegaard H., Sørum H. gyrA mutations in quinolone-resistant isolates of the fish pathogen Aeromonas salmonicida. Antimicrob Agents Chemother. 1994 Oct;38(10):2460–2464. doi: 10.1128/aac.38.10.2460. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. 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]
  21. Parales R. E., Harwood C. S. Nucleotide sequence of the gyrB gene of Pseudomonas putida. Nucleic Acids Res. 1990 Oct 11;18(19):5880–5880. doi: 10.1093/nar/18.19.5880. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Rahman M., Mauff G., Levy J., Couturier M., Pulverer G., Glasdorff N., Butzler J. P. Detection of 4-quinolone resistance mutation in gyrA gene of Shigella dysenteriae type 1 by PCR. Antimicrob Agents Chemother. 1994 Oct;38(10):2488–2491. doi: 10.1128/aac.38.10.2488. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Reece R. J., Maxwell A. DNA gyrase: structure and function. Crit Rev Biochem Mol Biol. 1991;26(3-4):335–375. doi: 10.3109/10409239109114072. [DOI] [PubMed] [Google Scholar]
  24. Reece R. J., Maxwell A. The C-terminal domain of the Escherichia coli DNA gyrase A subunit is a DNA-binding protein. Nucleic Acids Res. 1991 Apr 11;19(7):1399–1405. doi: 10.1093/nar/19.7.1399. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. 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]
  26. Reyna F., Huesca M., González V., Fuchs L. Y. Salmonella typhimurium gyrA mutations associated with fluoroquinolone resistance. Antimicrob Agents Chemother. 1995 Jul;39(7):1621–1623. doi: 10.1128/aac.39.7.1621. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Sreedharan S., Peterson L. R., Fisher L. M. Ciprofloxacin resistance in coagulase-positive and -negative staphylococci: role of mutations at serine 84 in the DNA gyrase A protein of Staphylococcus aureus and Staphylococcus epidermidis. Antimicrob Agents Chemother. 1991 Oct;35(10):2151–2154. doi: 10.1128/aac.35.10.2151. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Swanberg S. L., Wang J. C. Cloning and sequencing of the Escherichia coli gyrA gene coding for the A subunit of DNA gyrase. J Mol Biol. 1987 Oct 20;197(4):729–736. doi: 10.1016/0022-2836(87)90479-7. [DOI] [PubMed] [Google Scholar]
  29. 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]
  30. Vila J., Ruiz J., Goñi P., Marcos A., Jimenez de Anta T. Mutation in the gyrA gene of quinolone-resistant clinical isolates of Acinetobacter baumannii. Antimicrob Agents Chemother. 1995 May;39(5):1201–1203. doi: 10.1128/aac.39.5.1201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. 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]
  32. Whitby P. W., Landon M., Coleman G. The cloning and nucleotide sequence of the serine protease gene (aspA) of Aeromonas salmonicida ssp. salmonicida. FEMS Microbiol Lett. 1992 Nov 15;78(1):65–71. doi: 10.1016/0378-1097(92)90289-z. [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]

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

RESOURCES