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PMID: 1656457 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

An unusual mechanism for resistance to the antibiotic coumermycin A1.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 88 ·No. 19 ·1991-10-01 ·Pages 8860-4

del Castillo I, Vizán JL, Rodríguez-Sáinz MC, Moreno F

Abstract

Bacterial DNA gyrases are type II topoisomerases made up of two A subunits and two B subunits. Coumarins are carbohydrate-containing antibiotics that inhibit topoisomerases II by competing with ATP for binding to the enzymes. High resistance to coumarins is produced in bacterial species by mutations in gyrB, the gene encoding subunit B. We have found an unusual mechanism of resistance to coumarins in Escherichia coli. This mechanism is exhibited by cells containing the wild-type gyrB, or its 5' half, in high copy number. Since homologous mutant gyrB (coumermycin resistant) truncated genes did not confer drug resistance at all under the same conditions, we propose that this mechanism of resistance is due to drug sequestration by the overproduced wild-type GyrB polypeptides. A corollary of this is that the amino half of GyrB is required and sufficient to fashion the ATP-binding domain of DNA gyrase, a conclusion that was further supported by mapping three independent coumarin-resistant mutations at Arg-136 of GyrB. Just upstream of this residue there is a glycine-rich sequence highly conserved in all topoisomerases II, which seems to be a good candidate for the actual ATP-binding site.

Related Genes
gyrB
MeSH Terms
Amino Acid Sequence Aminocoumarins Cloning, Molecular Codon Coumarins/pharmacology DNA Mutational Analysis DNA Topoisomerases, Type II/physiology DNA, Bacterial/genetics Drug Resistance, Microbial Escherichia coli/drug effects Gene Expression Genes, Bacterial Molecular Sequence Data Plasmids Restriction Mapping
Chemicals
Aminocoumarins Codon Coumarins DNA, Bacterial DNA Topoisomerases, Type II coumermycin
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
del Castillo I
Unidad de Genética Molecular, Hospital Ramón y Cajal, Madrid, Spain.
Vizán J L
Rodríguez-Sáinz M C
Moreno F
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.88.19.8860
Published
1991-10-01
Pages
8860-4
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC52610
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.88.19.8860 PMC Full Text View journal details
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