Miconazole induces changes in actin cytoskeleton prior to reactive oxygen species induction in yeast.
The antifungal compound miconazole inhibits ergosterol biosynthesis and induces reactive oxygen species (ROS) in susceptible yeast species. To further uncover the mechanism of miconazole antifungal action and tolerance mechanisms, we screened the complete set of haploid Saccharomyces cerevisiae gene deletion mutants for mutants with an altered miconazole sensitivity phenotype. We identified 29 S. cerevisiae genes, which when deleted conferred at least 4-fold hypersensitivity to miconazole. Major functional groups encode proteins involved in tryptophan biosynthesis, membrane trafficking including endocytosis, regulation of actin cytoskeleton, and gene expression. With respect to the antifungal activity of miconazole, we demonstrate an antagonism with tryptophan and a synergy with a yeast endocytosis inhibitor. Because actin dynamics and induction of ROS are linked in yeast, we further focused on miconazole-mediated changes in actin cytoskeleton organization. In this respect, we demonstrate that miconazole induces changes in the actin cytoskeleton, indicative of increased filament stability, prior to ROS induction. These data provide novel mechanistic insights in the mode of action of a ROS-inducing azole.
Thevissen
Karin
K
Centre of Microbial and Plant Genetics, Katholieke Universiteit Leuven, Heverlee, Belgium.
Ayscough
Kathryn R
KR
Aerts
An M
AM
Du
Wei
W
De Brucker
Katrijn
K
Meert
Els M K
EM
Ausma
Jannie
J
Borgers
Marcel
M
Cammue
Bruno P A
BP
François
Isabelle E J A
IE
D000199:Actins / Q000187:drug effects* / Q000378:metabolism*; D003599:Cytoskeleton / Q000187:drug effects / Q000648:ultrastructure*; D004271:DNA, Fungal / Q000235:genetics; D008825:Miconazole / Q000494:pharmacology*; D016296:Mutagenesis; D010649:Phenylalanine / Q000494:pharmacology; D017382:Reactive Oxygen Species / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D017384:Sequence Deletion; D014364:Tryptophan / Q000494:pharmacology; D014443:Tyrosine / Q000494:pharmacology
D000199:Actins; D004271:DNA, Fungal; D017382:Reactive Oxygen Species; D014443:Tyrosine; D010649:Phenylalanine; D008825:Miconazole; D014364:Tryptophan