Antagonism between two mechanisms of antifungal drug resistance.
This study tested for interaction between two independently evolved mechanisms of fluconazole resistance in Saccharomyces cerevisiae. One set of strains was from a 400-generation evolution experiment, during which the concentration of fluconazole was increased from 16 to 256 microg/ml in four increments. At 100 generations, populations became fixed for resistance mutations in either of two transcriptional regulators, PDR1 or PDR3. At 400 generations, replicate populations became fixed for another resistance mutation in UNK1, an unmapped gene further increasing resistance. Another genotype used in this study came from a population placed initially in 128 microg/ml of fluconazole; this environment selects for resistance through loss of function at ERG3, resulting in altered sterol metabolism. Mutant strains carrying PDR1(r) or PDR3(r) were crossed with the erg3(r) mutant strain, and the doubly mutant, haploid offspring were identified. The double-mutant strains grew less well than the parent strains at all concentrations of fluconazole tested. In genome-wide assays of gene expression, several ABC transporter genes that were overexpressed in one parent and several ERG genes that were overexpressed in the other parent were also overexpressed in the double mutants. Of the 43 genes that were consistently overexpressed in the PDR1(r) parents at generation 100, however, 31 were not consistently overexpressed in the double mutants. Of these 31 genes, 30 were also not consistently overexpressed after a further 300 generations of evolution in the PDR1(r) parent populations. The two independently evolved mechanisms of fluconazole resistance are strongly antagonistic to one another.
Anderson
James B
JB
Department of Biology, University of Toronto, 3359 Mississauga Road North, Mississauga, Ontario L5L 1C6, Canada. janderso@utm.utoronto.ca
Ricker
Nicole
N
Sirjusingh
Caroline
C
D018528:ATP-Binding Cassette Transporters / Q000187:drug effects / Q000235:genetics; D000935:Antifungal Agents / Q000494:pharmacology; D003513:Cycloheximide / Q000494:pharmacology; D004268:DNA-Binding Proteins / Q000187:drug effects / Q000235:genetics; D019020:Directed Molecular Evolution; D025141:Drug Resistance, Fungal / Q000235:genetics* / Q000502:physiology; D051776:Early Growth Response Protein 3 / Q000187:drug effects / Q000235:genetics; D015725:Fluconazole / Q000494:pharmacology*; D015966:Gene Expression Regulation, Fungal / Q000187:drug effects; D008826:Microbial Sensitivity Tests; D009154:Mutation; D012235:Rhodamines / Q000494:pharmacology; D012441:Saccharomyces cerevisiae / Q000145:classification / Q000187:drug effects* / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins / Q000187:drug effects / Q000235:genetics; D015534:Trans-Activators / Q000187:drug effects / Q000235:genetics; D014157:Transcription Factors
D018528:ATP-Binding Cassette Transporters; D000935:Antifungal Agents; D004268:DNA-Binding Proteins; C085480:PDR1 protein, S cerevisiae; C092438:PDR5 protein, S cerevisiae; D012235:Rhodamines; D029701:Saccharomyces cerevisiae Proteins; D015534:Trans-Activators; D014157:Transcription Factors; C026188:rhodamine 6G; D051776:Early Growth Response Protein 3; D015725:Fluconazole; D003513:Cycloheximide