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PMID: 8083161 Published · ppublish English Journal Article

A Saccharomyces cerevisiae mutant with echinocandin-resistant 1,3-beta-D-glucan synthase.

Journal of bacteriology ·Vol. 176 ·No. 18 ·1994-09-00 ·Pages 5686-96

Douglas CM, Marrinan JA, Li W, Kurtz MB

Abstract

A novel, potent, semisynthetic pneumocandin, L-733,560, was used to isolate a resistant mutant in Saccharomyces cerevisiae. This compound, like other pneumocandins and echinocandins, inhibits 1,3-beta-D-glucan synthase from Candida albicans (F.A. Bouffard, R.A. Zambias, J. F. Dropinski, J.M. Balkovec, M.L. Hammond, G.K. Abruzzo, K.F. Bartizal, J.A. Marrinan, M. B. Kurtz, D.C. McFadden, K.H. Nollstadt, M.A. Powles, and D.M. Schmatz, J. Med. Chem. 37:222-225, 1994). Glucan synthesis catalyzed by a crude membrane fraction prepared from the S. cerevisiae mutant R560-1C was resistant to inhibition by L-733,560. The nearly 50-fold increase in the 50% inhibitory concentration against glucan synthase was commensurate with the increase in whole-cell resistance. R560-1C was cross-resistant to other inhibitors of C. albicans 1,3-beta-D-glucan synthase (aculeacin A, dihydropapulacandin, and others) but not to compounds with different modes of action. Genetic analysis revealed that enzyme and whole-cell pneumocandin resistance was due to a single mutant gene, designated etg1-1 (echinocandin target gene 1), which was semidominant in heterozygous diploids. The etg1-1 mutation did not confer enhanced ability to metabolize L-733,560 and had no effect on the membrane-bound enzymes chitin synthase I and squalene synthase. Alkali-soluble beta-glucan synthesized by crude microsomes from R560-1C was indistinguishable from the wild-type product. 1,3-beta-D-Glucan synthase activity from R560-1C was fractionated with NaCl and Tergitol NP-40; reconstitution with fractions from wild-type membranes revealed that drug resistance is associated with the insoluble membrane fraction. We propose that the etg1-1 mutant gene encodes a subunit of the 1,3-beta-D-glucan synthase complex.

Related Genes
etg1-1
MeSH Terms
Anti-Bacterial Agents/pharmacology Antifungal Agents/pharmacology Cell Membrane/chemistry Chitin Synthase/analysis Crosses, Genetic Drug Resistance, Microbial Farnesyl-Diphosphate Farnesyltransferase/analysis Genes, Fungal/genetics Glucosyltransferases/antagonists & inhibitors,biosynthesis,genetics,metabolism Kinetics Membrane Proteins Microsomes/enzymology Mutation/physiology Peptides Saccharomyces cerevisiae/drug effects,enzymology,genetics Schizosaccharomyces pombe Proteins
Chemicals
Anti-Bacterial Agents Antifungal Agents L 733560 Membrane Proteins Peptides Schizosaccharomyces pombe Proteins Glucosyltransferases Chitin Synthase 1,3-beta-glucan synthase Farnesyl-Diphosphate Farnesyltransferase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Douglas C M
Merck Research Laboratories, Rahway, New Jersey 07065.
Marrinan J A
Li W
Kurtz M B
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.176.18.5686-5696.1994
Published
1994-09-00
Pages
5686-96
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC196772
Subset
IM
External Links
PubMed source DOI: 10.1128/jb.176.18.5686-5696.1994 PMC Full Text View journal details
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