title: Application of a novel microtitre plate-based assay for the discovery of new inhibitors of DNA gyrase and DNA topoisomerase VI.
abstract: DNA topoisomerases are highly exploited targets for antimicrobial drugs. The spread of antibiotic resistance represents a significant threat to public health and necessitates the discovery of inhibitors that target topoisomerases in novel ways. However, the traditional assays for topoisomerase activity are not suitable for the high-throughput approaches necessary for drug discovery. In this study we validate a novel assay for screening topoisomerase inhibitors. A library of 960 compounds was screened against Escherichia coli DNA gyrase and archaeal Methanosarcina mazei DNA topoisomerase VI. Several novel inhibitors were identified for both enzymes, and subsequently characterised in vitro and in vivo. Inhibitors from the M. mazei topoisomerase VI screen were tested for their ability to inhibit Arabidopsis topoisomerase VI in planta. The data from this work present new options for antibiotic drug discovery and provide insight into the mechanism of topoisomerase VI.
authors.affiliation: Department of Biological Chemistry, John Innes Centre, Colney Lane, Norwich, United Kingdom.
authors.forename: James A
authors.initials: JA
authors.lastname: Taylor
authors.forename: Lesley A
authors.initials: LA
authors.lastname: Mitchenall
authors.forename: Martin
authors.initials: M
authors.lastname: Rejzek
authors.forename: Robert A
authors.initials: RA
authors.lastname: Field
authors.forename: Anthony
authors.initials: A
authors.lastname: Maxwell
mesh_terms: D000880:Anthraquinones / Q000494:pharmacology; D017360:Arabidopsis / Q000187:drug effects / Q000201:enzymology / Q000254:growth & development; D019843:Archaeal Proteins / Q000037:antagonists & inhibitors*; D004250:DNA Topoisomerases, Type II; D004353:Drug Evaluation, Preclinical / Q000295:instrumentation / Q000379:methods*; D004926:Escherichia coli / Q000201:enzymology; D006604:Hexylresorcinol / Q000494:pharmacology; D017020:Methanosarcina / Q000201:enzymology; D008942:Mitoxantrone / Q000494:pharmacology; D011796:Quinacrine / Q000494:pharmacology; D016944:Sulfolobus / Q000201:enzymology; D013498:Suramin / Q000494:pharmacology; D059005:Topoisomerase II Inhibitors; D059003:Topoisomerase Inhibitors / Q000494:pharmacology*
chemical_list: D000880:Anthraquinones; D019843:Archaeal Proteins; D059005:Topoisomerase II Inhibitors; D059003:Topoisomerase Inhibitors; D013498:Suramin; D008942:Mitoxantrone; C105141:DNA topoisomerase VI; D004250:DNA Topoisomerases, Type II; D011796:Quinacrine; C511975:purpurin anthraquinone; D006604:Hexylresorcinol