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PMID: 1850970 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Novel quinolone resistance mutations of the Escherichia coli DNA gyrase A protein: enzymatic analysis of the mutant proteins.

Antimicrobial agents and chemotherapy ·Vol. 35 ·No. 2 ·1991-02-00 ·Pages 335-40

Hallett P, Maxwell A

Abstract

Using the techniques of gap misrepair mutagenesis and site-directed mutagenesis, we have generated two novel quinolone resistance mutations of the Escherichia coli DNA gyrase A protein. DNA sequencing showed these mutations to be Ser-83----Ala and Gln-106----Arg. The mutant proteins were overproduced and purified, and their enzymatic properties were analyzed and compared with those of the wild-type enzyme. With ciprofloxacin and other quinolones, the inhibition of DNA supercoiling, relaxation, and decatenation and the induction of DNA cleavage were investigated for both wild-type and mutant enzymes. In each assay, the mutant enzymes were found to require approximately 10 times more drug to inhibit the reaction or induce cleavage than was the wild-type enzyme. However, the Ca2(+)-directed DNA cleavage reaction was indistinguishable for wild-type and mutant gyrases. We discuss models for the gyrase-mediated bactericidal effects of quinolone drugs.

MeSH Terms
4-Quinolones Anti-Infective Agents/pharmacology DNA Repair DNA Topoisomerases, Type II/genetics,metabolism Drug Resistance, Microbial Escherichia coli/drug effects,enzymology,genetics Mutagenesis Plasmids Protein Denaturation
Chemicals
4-Quinolones Anti-Infective Agents DNA Topoisomerases, Type II
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Hallett P
Department of Biochemistry, University of Leicester, United Kingdom.
Maxwell A
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44 references, click to expand
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Article Info
Journal
Antimicrobial agents and chemotherapy
Abbr.
Antimicrob Agents Chemother
ISSN
0066-4804
DOI
10.1128/AAC.35.2.335
Published
1991-02-00
Pages
335-40
Language
English
Region
United States
NLM ID
0315061
PMCID
PMC245001
Subset
IM
External Links
PubMed source DOI: 10.1128/AAC.35.2.335 PMC Full Text View journal details
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