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

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.

Use of a naturally occurring codon bias for identifying topoisomerase mutations in ciprofloxacin resistant <i>Escherichia coli</i> using PCR and future prospects with other bacterial genera: A pilot study

“…We observed similar mutations (Ser to Leu and Asp to Asn) in the both the strains that possessed a double mutation when subjected to sequencing. This observation is also similar with earlier reports [6,7]. The enteropathogenic E. coli (EPEC) reference strain E2348/69 (O127: H6) was found to be ciprofloxacin sensitive in our earlier work [10].…”

“…Mutations at ser83 of gyrA is the most frequent in Escherichia coli (E. coli) clinical isolates [5]. Mutation of ser83 to leucine (Leu) or tryptophan (Trp) confers high level resistance to quinolones while ser83 to Alanine (Ala) confers low level resistance [6]. Similarly, aspartate at residue 87 (asp87) is another important target for quinolone drugs.…”

In Vitro Activity of Five Quinolones and Analysis of the Quinolone Resistance-Determining Regions of gyrA , gyrB , parC , and parE in Ureaplasma parvum and Ureaplasma urealyticum Clinical Isolates from Perinatal Patients in Japan

“…with a high MIC against quinolones are already widely distributed among females of reproductive age in Japan, which may complicate the eradication of ureaplasmas from the urogenital tract even in nonpregnant reproductive women, underscoring the importance of keeping quinolone treatment of adult NGU and respiratory infections under surveillance. The S83L and S83W mutations were detected in the gyrA gene of E. coli and Staphylococcus pseudintermedius (S84W) (29,30). A GyrA A87V/S88P (corresponding to positions 83 and 84, respectively, in Escherichia coli) double mutant was recently reported in multidrug-resistant Ochrobactrum intermedium (31).…”

Quinolonas: Perspectivas actuales y mecanismos de resistencia

“…Se dan por errores de transcripción durante la replicación cromosómica y ocurren en rangos tan altos como 1 en 10 6 a 10 9 en bacterias silvestres 34 . Las mutaciones en gyrA, el gen que codifica las subunidades A de ADN girasa, conforman el mecanismo que con mayor frecuencia se encuentra involucrado en la resistencia a quinolonas entre las bacterias gramnegativas 35 .…”