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Antimicrobial Agents and Chemotherapy, September 1998, p. 2304-2311, Vol. 42, No. 9
0066-4804/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Alterations in Topoisomerase IV and DNA Gyrase in Quinolone-Resistant Mutants of Mycoplasma hominis Obtained In Vitro

Cécile M. Bébéar,1,2,* Hélène Renaudin,1 Alain Charron,1 Joseph M. Bové,2 Christiane Bébéar,1 and Joel Renaudin2

Laboratoire de Bactériologie, Université Bordeaux 2, 33076 Bordeaux cedex,1 and Laboratoire de Biologie Cellulaire et Moléculaire, Institut National de la Recherche Agronomique, 33883 Villenave d'Ornon cedex,2 France

Received 3 November 1997/Returned for modification 23 January 1998/Accepted 24 March 1998

Mycoplasma hominis mutants were selected stepwise for resistance to ofloxacin and sparfloxacin, and their gyrA, gyrB, parC, and parE quinolone resistance-determining regions were characterized. For ofloxacin, four rounds of selection yielded six first-, six second-, five third-, and two fourth-step mutants. The first-step mutants harbored a single Asp426right-arrowAsn substitution in ParE. GyrA changes (Ser83right-arrowLeu or Trp) were found only from the third round of selection. With sparfloxacin, three rounds of selection generated 4 first-, 7 second-, and 10 third-step mutants. In contrast to ofloxacin resistance, GyrA mutations (Ser83right-arrowLeu or Ser84right-arrowTrp) were detected in the first-step mutants prior to ParC changes (Glu84right-arrowLys), which appeared only after the second round of selection. Further analysis of eight multistep-selected mutants of M. hominis that were previously described (2) revealed that they carried mutations in ParE (Asp426right-arrowAsn), GyrA (Ser83right-arrowLeu) and ParE (Asp426right-arrowAsn), GyrA (Ser83right-arrowLeu) and ParC (Ser80right-arrowIle), or ParC (Ser80right-arrowIle) alone, depending on the fluoroquinolone used for selection, i.e., ciprofloxacin, norfloxacin, ofloxacin, or pefloxacin, respectively. These data indicate that in M. hominis DNA gyrase is the primary target of sparfloxacin whereas topoisomerase IV is the primary target of pefloxacin, ofloxacin, and ciprofloxacin.

* Corresponding author. Mailing address: Laboratoire de Bactériologie, Université Bordeaux 2, 146 rue Léo Saignat, 33076 Bordeaux cedex, France. Phone: (33) 5.57.57.16.25. Fax: (33) 5.56.79.56.11. E-mail: cecile.bebear{at}labbebear.u-bordeaux2.fr.

Antimicrobial Agents and Chemotherapy, September 1998, p. 2304-2311, Vol. 42, No. 9
0066-4804/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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