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Antimicrobial Agents and Chemotherapy, December 2004, p. 4495-4504, Vol. 48, No. 12
0066-4804/04/$08.00+0     DOI: 10.1128/AAC.48.12.4495-4504.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

A Mutation in Escherichia coli DNA Gyrase Conferring Quinolone Resistance Results in Sensitivity to Drugs Targeting Eukaryotic Topoisomerase II

Thomas Gruger,1 John L. Nitiss,2 Anthony Maxwell,3 E. Lynn Zechiedrich,4 Peter Heisig,1 Siegfried Seeber,5 Yves Pommier,6 and Dirk Strumberg6*

Department of Pharmaceutical Biology & Microbiology, Institute of Pharmacy, University of Hamburg, Hamburg, Germany,1 Department of Molecular Pharmacology, St. Jude Children's Research Hospital, Memphis, Tennessee,2 Department of Biochemistry, University of Leicester, Leicester, United Kingdom,3 Department of Molecular Virology & Microbiology, Baylor College of Medicine, Houston, Texas,4 Department of Internal Medicine and Medical Oncology, West German Cancer Center, University Medical School of Essen, Essen, Germany,5 Laboratory of Molecular Pharmacology, Division of Basic Sciences, National Cancer Institute, National Institutes of Health, Bethesda, Maryland6

Received 1 March 2004/ Returned for modification 11 July 2004/ Accepted 19 August 2004

Fluoroquinolones are broad-spectrum antimicrobial agents that target type II topoisomerases. Many fluoroquinolones are highly specific for bacterial type II topoisomerases and act against both DNA gyrase and topoisomerase IV. In Escherichia coli, mutations causing quinolone resistance are often found in the gene that encodes the A subunit of DNA gyrase. One common site for resistance-conferring mutations alters Ser83, and mutations to Leu or Trp result in high levels of resistance to fluoroquinolones. In the present study we demonstrate that the mutation of Ser83 to Trp in DNA gyrase (GyrS83W) also results in sensitivity to agents that are potent inhibitors of eukaryotic topoisomerase II but that are normally inactive against prokaryotic enzymes. Epipodophyllotoxins, such as etoposide, teniposide and amino-azatoxin, inhibited the DNA supercoiling activity of GyrS83W, and the enzyme caused elevated levels of DNA cleavage in the presence of these agents. The DNA sequence preference for GyrS83W-induced cleavage sites in the presence of etoposide was similar to that seen with eukaryotic type II topoisomerases. Introduction of the GyrS83W mutation in E. coli strain RFM443-242 by site-directed mutagenesis sensitized it to epipodophyllotoxins and amino-azatoxin. Our results demonstrate that sensitivity to agents that target topoisomerase II is conserved between prokaryotic and eukaryotic enzymes, suggesting that drug interaction domains are also well conserved and likely occur in domains important for the biochemical activities of the enzymes.

* Corresponding author. Present address: Department of Internal Medicine (Cancer Research), University Medical School of Essen, Hufelandstr. 55, 45122 Essen, Germany. Phone: 49-(0)201-723-2027. Fax: 49-(0)201-723-5988. E-mail: dirk.strumberg{at}uni-essen.de.

Antimicrobial Agents and Chemotherapy, December 2004, p. 4495-4504, Vol. 48, No. 12
0066-4804/04/$08.00+0     DOI: 10.1128/AAC.48.12.4495-4504.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Heisig, P. (2009). Type II topoisomerases--inhibitors, repair mechanisms and mutations. Mutagenesis 24: 465-469 [Abstract] [Full Text]  
  • Duca, M., Guianvarc'h, D., Oussedik, K., Halby, L., Garbesi, A., Dauzonne, D., Monneret, C., Osheroff, N., Giovannangeli, C., Arimondo, P. B. (2006). Molecular basis of the targeting of topoisomerase II-mediated DNA cleavage by VP16 derivatives conjugated to triplex-forming oligonucleotides.. Nucleic Acids Res 34: 1900-1911 [Abstract] [Full Text]  


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