We identified YPT31, which is involved in Golgi traffic, as a clotrimazole (CTZ)-resistance gene in a multicopy library screen. Multicopies of the YPT31 homolog YPT32 also conferred resistance to CTZ, and single disruption of YPT31 or YPT32 resulted in sensitivity to CTZ. Pdr5p, an ATP-binding cassette (ABC) transporter at the plasma membrane, was the most important factor for mediating basal resistance to CTZ, suggesting that Ypt31p and Ypt32p might be involved in the trafficking of Pdr5p to the plasma membrane. However, the activity of Pdr5p was independent of YPT31 or YPT32, and multicopies of YPT31 or YPT32 still conferred resistance to CTZ in pdr5 cells. To elucidate the roles of YPT31 and YPT32 in CTZ resistance, we analyzed mutants of 11 genes that are involved in the following vesicular trafficking: Golgi traffic (kes1, trs33, trs65, gyp1, trs85, and gyp2), vacuole inheritance (ypt7), endocytosis (rcy1 and ypt51) and exocytosis (msb3 and msb4). All of the mutant cells except ypt51, msb3 and msb4 were sensitive to CTZ, indicating that vacuoles were involved in CTZ resistance, since vacuole formation requires proper Golgi-trafficking and endocytosis. Microscopic analysis showed abnormal vacuoles in ypt31 cells. Multicopies of YPT31 or YPT32 conferred resistance to CTZ in AD1-8 cells, which are defective in seven major drug transporters, and in pdr5 ypt7 cells, but not in ypt7 or AD1-8-7 (AD1-8/ypt7) cells. These results indicated that Ypt31p and Ypt32p played minor but compensatory roles in cellular resistance to CTZ through vacuoles and specific ABC transporter(s) other than Pdr5p.
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Evidence ID | Analyze ID | Gene/Complex | Systematic Name/Complex Accession | Qualifier | Gene Ontology Term ID | Gene Ontology Term | Aspect | Annotation Extension | Evidence | Method | Source | Assigned On | Reference |
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Evidence ID | Analyze ID | Gene | Gene Systematic Name | Phenotype | Experiment Type | Experiment Type Category | Mutant Information | Strain Background | Chemical | Details | Reference |
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Evidence ID | Analyze ID | Gene | Gene Systematic Name | Disease Ontology Term | Disease Ontology Term ID | Qualifier | Evidence | Method | Source | Assigned On | Reference |
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Evidence ID | Analyze ID | Regulator | Regulator Systematic Name | Target | Target Systematic Name | Direction | Regulation of | Happens During | Regulator Type | Direction | Regulation Of | Happens During | Method | Evidence | Strain Background | Reference |
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Site | Modification | Modifier | Source | Reference |
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Evidence ID | Analyze ID | Interactor | Interactor Systematic Name | Interactor | Interactor Systematic Name | Allele | Assay | Annotation | Action | Phenotype | SGA score | P-value | Source | Reference | Note |
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Evidence ID | Analyze ID | Interactor | Interactor Systematic Name | Interactor | Interactor Systematic Name | Assay | Annotation | Action | Modification | Source | Reference | Note |
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Complement ID | Locus ID | Gene | Species | Gene ID | Strain background | Direction | Details | Source | Reference |
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Evidence ID | Analyze ID | Dataset | Description | Keywords | Number of Conditions | Reference |
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