Quinolone resistance mutations in the DNA gyrase gyrA and gyrB genes of Staphylococcus aureus
- 1 September 1994
- journal article
- research article
- Published by American Society for Microbiology in Antimicrobial Agents and Chemotherapy
- Vol. 38 (9), 2014-2023
- https://doi.org/10.1128/aac.38.9.2014
Abstract
A 6.4-kb DNA fragment containing the DNA gyrase gyrA and gyrB genes was cloned and sequenced from the quinolone-susceptible Staphylococcus aureus type strain ATCC 12600. An expression plasmid was constructed by inserting the cloned genes into the Escherichia coli-S. aureus shuttle vector pAT19, and deletion plasmids carrying only functional gyrA and gyrB genes were derived from this plasmid. An efficient transformation system for S. aureus RN4220 was established by using these plasmids. Quinolone-resistant mutants of S. aureus RN4220 were isolated by three-step selection with quinolones. The first- and second-step mutants were considered to be transport mutants, and the third-step mutants were divided into five groups with respect to their resistance patterns and transformation results with gyrA and gyrB genes. Sequencing analysis of the resulting mutant gyrase genes showed that they had the following point mutations: group 1, Ser-84 (TCA) to Leu (TTA) in GyrA; group 2, Ser-84 (TCA) to Ala (GCA), Ser-85 (TCT) to Pro (CCT), or Glu-88 (GAA) to Lys (AAA) in GyrA; group 3, Asp-437 (GAC) to Asn (AAC) in GyrB; group 4, Arg-458 (CGA) to Gln (CAA) in GyrB; and group 5, Ser-85 (TCT) to Pro (CCT) in GyrA and Asp-437 (GAC) to Asn (AAC) in GyrB. When the gyrA and/or gyrB mutants were transformed with the wild-type gyrA and/or gyrB plasmids, they became quinolone susceptible, but transformants with the plasmids having the same mutations on the gyrA and/or gyrB genes did not confer susceptibility. These results indicate that mutations in both gyrA and gyrB can be responsible for quinolone resistance in S. aureus.Keywords
This publication has 35 references indexed in Scilit:
- Antimicrobial activity of DU-6859, a new potent fluoroquinolone, against clinical isolatesAntimicrobial Agents and Chemotherapy, 1992
- Detection of gyrA gene mutations associated with ciprofloxacin resistance in methicillin-resistant Staphylococcus aureus: analysis by polymerase chain reaction and automated direct DNA sequencingAntimicrobial Agents and Chemotherapy, 1992
- gyrA Mutations in Ciprofloxacin-Resistant, Methicillin-Resistant Staphylococcus aureus from Indiana, Minnesota, and TennesseeThe Journal of Infectious Diseases, 1991
- Crystal structure of an N-terminal fragment of the DNA gyrase B proteinNature, 1991
- Shuttle vectors containing a multiple cloning site and a lacZα gene for conjugal transfer of DNA from Escherichia coli to Gram-positive bacteriaGene, 1991
- Pyridonecarboxylic acids as antibacterial agents. Part 14. Synthesis and structure-activity relationships of 5-substituted 6,8-difluoroquinolones, including sparfloxacin, a new quinolone antibacterial agent with improved potencyJournal of Medicinal Chemistry, 1990
- The Leucine Zipper: A Hypothetical Structure Common to a New Class of DNA Binding ProteinsScience, 1988
- The toxic shock syndrome exotoxin structural gene is not detectably transmitted by a prophageNature, 1983
- Structure-activity relationships of antibacterial 6,7- and 7,8-disubstituted 1-alkyl-1,4-dihydro-4-oxoquinoline-3-carboxylic acidsJournal of Medicinal Chemistry, 1980
- Quinoline antibacterial agents. Oxolinic acid and related compoundsJournal of Medicinal Chemistry, 1968