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Mitochondrial resistance to miconazole in Saccharomyces cerevisiae
Abstract:
One mutant of mitochondrial origin resistant to miconazole has been isolated and characterized in S. cerevisiae. The mutation is linked to the locus oli1, the structural gene for subunit 9 of ATPase on mitochondrial DNA. Miconazole inhibited the mitochondrial ATPase of the wild type while the enzyme of the resistant mutant was insensitive to this effect. Levels of ATP decreased to one-third of the control in the wild type in the presence of miconazole, while they were unaffected in the mutant.
Insights
A yeast mutant resistant to miconazole was identified. This resistance stems from a mutation in the mitochondrial gene for ATPase subunit 9, protecting cellular ATP levels from the drug.
Area of Science:
- Biochemistry
- Molecular Biology
- Yeast Genetics
Background:
- Mitochondrial ATPase is crucial for cellular energy production.
- Miconazole is an antifungal agent that can inhibit mitochondrial function.
- Genetic mutations can confer resistance to drugs affecting cellular processes.
Purpose of the Study:
- To isolate and characterize a miconazole-resistant mutant in Saccharomyces cerevisiae.
- To investigate the genetic basis of miconazole resistance in yeast mitochondria.
- To determine the effect of miconazole on mitochondrial ATPase activity and ATP levels in wild-type and mutant strains.
Main Methods:
- Isolation and characterization of a miconazole-resistant mutant.
- Genetic mapping of the resistance mutation to the oli1 locus on mitochondrial DNA.
- Enzyme assays measuring mitochondrial ATPase activity.
- Measurement of intracellular ATP levels.
Main Results:
- A single mitochondrial mutation conferring miconazole resistance was identified in S. cerevisiae.
- The mutation is located in the oli1 gene, encoding ATPase subunit 9.
- Miconazole inhibited wild-type mitochondrial ATPase but not the mutant enzyme.
- Miconazole treatment reduced ATP levels in wild-type cells but not in the resistant mutant.
Conclusions:
- The oli1 gene mutation confers resistance to miconazole by altering mitochondrial ATPase sensitivity.
- This mutation protects cellular ATP levels from miconazole-induced inhibition.
- Mitochondrial ATPase is a key target for miconazole's antifungal activity in S. cerevisiae.