TY - JOUR
T1 - Membrane rafts are involved in intracellular miconazole accumulation in yeast cells
AU - François, Isabelle E.J.A.
AU - Bink, Anna
AU - Vandercappellen, Jo
AU - Ayscough, Kathryn R.
AU - Toulmay, Alexandre
AU - Schneiter, Roger
AU - van Gyseghem, Elke
AU - Van den Mooter, Guy
AU - Borgers, Marcel
AU - Vandenbosch, Davy
AU - Coenye, Tom
AU - Cammue, Bruno P.A.
AU - Thevissen, Karin
PY - 2009/11/20
Y1 - 2009/11/20
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=70450235095&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=70450235095&partnerID=8YFLogxK
U2 - 10.1074/jbc.M109.014571
DO - 10.1074/jbc.M109.014571
M3 - Article
C2 - 19783660
AN - SCOPUS:70450235095
SN - 0021-9258
VL - 284
SP - 32680
EP - 32685
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 47
ER -