Disruption of the Saccharomyces cerevisiae gene for NADPH-cytochrome P450 reductase causes increased sensitivity to ketoconazole.
Strains of Saccharomyces cerevisiae deleted in the NADPH-cytochrome P450 reductase gene by transplacement are 200-fold more sensitive to ketoconazole, an inhibitor of the cytochrome P450 lanosterol 14 alpha-demethylase. Resistance is restored through complementation by the plasmid-borne wild type gene from either S. cerevisiae or Candida tropicalis. Neither Southern hybridization nor Western immunoblot techniques provided evidence for a second NADPH-cytochrome P450 reductase gene, suggesting that an alternate pathway may provide for the functions of this reductase in S. cerevisiae.
Sutter
T R
TR
Department of Environmental Health, University of Cincinnati College of Medicine, Ohio 45267-0524.
Loper
J C
JC
D015153:Blotting, Western; D002175:Candida / Q000235:genetics; D003577:Cytochrome P-450 Enzyme System; D004262:DNA Restriction Enzymes; D004271:DNA, Fungal / Q000302:isolation & purification; D004352:Drug Resistance, Microbial / Q000235:genetics; D005800:Genes, Fungal; D007654:Ketoconazole / Q000494:pharmacology*; D007930:Leucine / Q000502:physiology; D009251:NADPH-Ferrihemoprotein Reductase / Q000235:genetics*; D009693:Nucleic Acid Hybridization; D010088:Oxidoreductases / Q000037:antagonists & inhibitors; D010641:Phenotype; D010957:Plasmids; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development; D058886:Sterol 14-Demethylase; D014170:Transformation, Genetic
D004271:DNA, Fungal; D003577:Cytochrome P-450 Enzyme System; D010088:Oxidoreductases; D058886:Sterol 14-Demethylase; D009251:NADPH-Ferrihemoprotein Reductase; D004262:DNA Restriction Enzymes; D007930:Leucine; D007654:Ketoconazole