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

Antimicrobial Agents and Chemotherapy - Tập 48 Số 12 - Trang 4495-4504 - 2004
Thomas Grüger1, John L. Nitiss2, Anthony Maxwell3, Lynn Zechiedrich4, Peter Heisig1, S. Seeber5, Yves Pommier6, Dirk Strumberg6
1Department of Pharmaceutical Biology & Microbiology, Institute of Pharmacy, University of Hamburg, Hamburg, Germany
2Department of Molecular Pharmacology, St. Jude Children's Research Hospital, Memphis, Tennessee
3Department of Biochemistry, University of Leicester, Leicester, United Kingdom
4Department of Molecular Virology & Microbiology, Baylor College of Medicine, Houston, Texas
5Department of Internal Medicine and Medical Oncology, West German Cancer Center, University Medical School of Essen, Essen, Germany
6Laboratory of Molecular Pharmacology, Division of Basic Sciences, National Cancer Institute, National Institutes of Health, Bethesda, Maryland

Tóm tắt

ABSTRACT

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 Ser 83 , and mutations to Leu or Trp result in high levels of resistance to fluoroquinolones. In the present study we demonstrate that the mutation of Ser 83 to Trp in DNA gyrase (Gyr S83W ) 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 Gyr S83W , and the enzyme caused elevated levels of DNA cleavage in the presence of these agents. The DNA sequence preference for Gyr S83W -induced cleavage sites in the presence of etoposide was similar to that seen with eukaryotic type II topoisomerases. Introduction of the Gyr S83W 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.

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