title: Miconazole induces changes in actin cytoskeleton prior to reactive oxygen species induction in yeast.
abstract: 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.
authors.affiliation: Centre of Microbial and Plant Genetics, Katholieke Universiteit Leuven, Heverlee, Belgium.
authors.forename: Karin
authors.initials: K
authors.lastname: Thevissen
authors.forename: Kathryn R
authors.initials: KR
authors.lastname: Ayscough
authors.forename: An M
authors.initials: AM
authors.lastname: Aerts
authors.forename: Wei
authors.initials: W
authors.lastname: Du
authors.forename: Katrijn
authors.initials: K
authors.lastname: De Brucker
authors.forename: Els M K
authors.initials: EM
authors.lastname: Meert
authors.forename: Jannie
authors.initials: J
authors.lastname: Ausma
authors.forename: Marcel
authors.initials: M
authors.lastname: Borgers
authors.forename: Bruno P A
authors.initials: BP
authors.lastname: Cammue
authors.forename: Isabelle E J A
authors.initials: IE
authors.lastname: François
mesh_terms: 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
chemical_list: D000199:Actins; D004271:DNA, Fungal; D017382:Reactive Oxygen Species; D014443:Tyrosine; D010649:Phenylalanine; D008825:Miconazole; D014364:Tryptophan