Membrane rafts are involved in intracellular miconazole accumulation in yeast cells

François, Isabelle E. J. A.; Bink, Anna; Vandercappellen, Jo; Ayscough, Kathryn R.; Toulmay, Alexandre; Schneiter, Roger; Gyseghem, Elke van; Mooter, Guy Van den; Borgers, Marcel; Vandenbosch, Davy; Coenye, Tom; Cammue, Bruno P. A.; Thevissen, Karin

doi:10.1074/jbc.M109.014571

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Journal article

Membrane rafts are involved in intracellular miconazole accumulation in yeast cells

François, Isabelle E. J. A. Centre of Microbial and Plant Genetics, Katholieke Universiteit Leuven, Belgium
Bink, Anna Centre of Microbial and Plant Genetics, Katholieke Universiteit Leuven, Belgium
Vandercappellen, Jo Centre of Microbial and Plant Genetics, Katholieke Universiteit Leuven, Belgium
Ayscough, Kathryn R. Department of Molecular Biology and Biotechnology, University of Sheffield, UK
Toulmay, Alexandre Division of Biochemistry, University of Fribourg, Switzerland
Schneiter, Roger Division of Biochemistry, University of Fribourg, Switzerland
Gyseghem, Elke van Laboratory of Pharmaceutical Technology and Biopharmacy, Katholieke Universiteit Leuven, Belgium
Mooter, Guy Van den Laboratory of Pharmaceutical Technology and Biopharmacy, Katholieke Universiteit Leuven, Belgium
Borgers, Marcel Barrier Therapeutics Geel, Belgium
Vandenbosch, Davy Laboratorium voor Farmaceutische Microbiologie, Universiteit Gent, Belgium
Coenye, Tom Laboratorium voor Farmaceutische Microbiologie, Universiteit Gent, Belgium
Cammue, Bruno P. A. Centre of Microbial and Plant Genetics, Katholieke Universiteit Leuven, Belgium
Thevissen, Karin Centre of Microbial and Plant Genetics, Katholieke Universiteit Leuven, Belgium

20.11.2009

Published in:

The Journal of Biological Chemistry. - 2009, vol. 284, p. 32680-32685

English Azoles inhibit ergosterol biosynthesis, resulting in ergosterol depletion and accumulation of toxic 14α-methylated sterols in membranes of susceptible yeast. We demonstrated previously that miconazole induces actin cytoskeleton stabilization in Saccharomyces cerevisiae prior to induction of reactive oxygen species, pointing to an ancillary mode of action. Using a genome-wide agar-based screening, we demonstrate in this study that S. cerevisiae mutants affected in sphingolipid and ergosterol biosynthesis, namely ipt1, sur1, skn1, and erg3 deletion mutants, are miconazole-resistant, suggesting an involvement of membrane rafts in its mode of action. This is supported by the antagonizing effect of membrane raft-disturbing compounds on miconazole antifungal activity as well as on miconazole-induced actin cytoskeleton stabilization and reactive oxygen species accumulation. These antagonizing effects point to a primary role for membrane rafts in miconazole antifungal activity. We further show that this primary role of membrane rafts in miconazole action consists of mediating intracellular accumulation of miconazole in yeast cells.

Faculty

Faculté des sciences et de médecine

Department

Département de Biologie

Language