Topoisomerase IV-quinolone interactions are mediated through a water-metal ion bridge: mechanistic basis of quinolone resistance

Katie J Aldred; Sylvia A McPherson; Charles L Turnbough Jr; Robert J Kerns; Neil Osheroff

doi:10.1093/nar/gkt124

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Topoisomerase IV-quinolone interactions are mediated through a water-metal ion bridge: mechanistic basis of quinolone resistance

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Topoisomerase IV-quinolone interactions are mediated through a water-metal ion bridge: mechanistic basis of quinolone resistance

Katie J Aldred, Sylvia A McPherson, Charles L Turnbough Jr, Robert J Kerns and Neil Osheroff

Nucleic acids research, Vol.41(8), pp.4628-4639

04/2013

DOI: 10.1093/nar/gkt124

PMCID: PMC3632122

PMID: 23460203

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Abstract

Although quinolones are the most commonly prescribed antibacterials, their use is threatened by an increasing prevalence of resistance. The most common causes of quinolone resistance are mutations of a specific serine or acidic residue in the A subunit of gyrase or topoisomerase IV. These amino acids are proposed to serve as a critical enzyme-quinolone interaction site by anchoring a water-metal ion bridge that coordinates drug binding. To probe the role of the proposed water-metal ion bridge, we characterized wild-type, GrlA(E85K), GrlA(S81F/E85K), GrlA(E85A), GrlA(S81F/E85A) and GrlA(S81F) Bacillus anthracis topoisomerase IV, their sensitivity to quinolones and related drugs and their use of metal ions. Mutations increased the Mg(2+) concentration required to produce maximal quinolone-induced DNA cleavage and restricted the divalent metal ions that could support quinolone activity. Individual mutation of Ser81 or Glu85 partially disrupted bridge function, whereas simultaneous mutation of both residues abrogated protein-quinolone interactions. Results provide functional evidence for the existence of the water-metal ion bridge, confirm that the serine and glutamic acid residues anchor the bridge, demonstrate that the bridge is the primary conduit for interactions between clinically relevant quinolones and topoisomerase IV and provide a likely mechanism for the most common causes of quinolone resistance.

Drug Resistance

Mutation

Anti-Bacterial Agents - chemistry

Anti-Bacterial Agents - pharmacology

Bacillus anthracis - enzymology

Cations, Divalent - chemistry

Ciprofloxacin - chemistry

Ciprofloxacin - pharmacology

DNA - metabolism

DNA Cleavage - drug effects

DNA Topoisomerase IV - chemistry

DNA Topoisomerase IV - drug effects

DNA Topoisomerase IV - genetics

DNA Topoisomerase IV - metabolism

Magnesium - chemistry

Metals - chemistry

Quinazolinones - chemistry

Quinazolinones - pharmacology

Quinolones - chemistry

Water - chemistry

Details

Title: Subtitle: Topoisomerase IV-quinolone interactions are mediated through a water-metal ion bridge: mechanistic basis of quinolone resistance
Creators: Katie J Aldred - Vanderbilt University
Sylvia A McPherson - Vanderbilt University
Charles L Turnbough Jr
Robert J Kerns
Neil Osheroff - Vanderbilt University
Resource Type: Journal article
Publication Details: Nucleic acids research, Vol.41(8), pp.4628-4639
DOI: 10.1093/nar/gkt124
PMID: 23460203
PMCID: PMC3632122
NLM abbreviation: Nucleic Acids Res
ISSN: 0305-1048
eISSN: 1362-4962
Grant note: T32 CA09582 / NCI NIH HHS R01 AI087671 / NIAID NIH HHS GM033944 / NIGMS NIH HHS AI81775 / NIAID NIH HHS T32 CA009582 / NCI NIH HHS GM33944 / NIGMS NIH HHS AI87671 / NIAID NIH HHS
Language: English
Date published: 04/2013
Academic Unit: Pharmaceutical Sciences and Experimental Therapeutics; Medicinal and Natural Products Chemistry
Record Identifier: 9984365879202771

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