Project description:Time-course analyses of the modifications of the yeast gene expression program which immediately follows addition of the antimitotic drug benomyl. Parallel experiments were conducted in different genetic contexts using strains deleted in the Yap1 and Pdr1 genes. Keywords: other
Project description:Time-course analyses of the modifications of the yeast gene expression program which immediately follows addition of the antimitotic drug benomyl. Parallel experiments were conducted in different genetic contexts using strains deleted in the Yap1 and Pdr1 genes.
Project description:Yeast is a powerful model system for studying the action of small molecule therapeutics. An important limitation has been low efficacy of many small molecules in yeast due to limited intracellular drug accumulation. We used the DNA binding domain of the pleiotropic drug resistance regulator Pdr1 fused in-frame to transcription repressors to repress Pdr1 regulated genes. Expression of these regulators conferred dominant enhancement of drug sensitivity and led to greatly diminished levels of Pdr1p regulated transcripts, including the yeast p-glycoprotein homologue Pdr5. Enhanced sensitivity was seen for a wide range of small molecules. Biochemical measurements demonstrated enhanced accumulation of rhodamine in yeast cells carrying the chimeras. These repressors of Pdr1p regulated transcripts can be introduced into large collections of strains such as the S. cerevisiae deletion set, and enhance the utility of yeast for studying drug action and for mechanism-based drug discovery. Keywords: Comparison of genetic variants
Project description:Growth assay in the presence of a toxic chemical (sr7575) that uses the barcoded collections of yeast gene deletions (haploid, diploid, DamP) to identify deletion strains that are hypersensitive to the drug.
Project description:Yap1 targets under normal and cobalt surplus growth conditions. Yeast strains (wild-type and yap1 mutant, BY4742 background) were grown until early log-phase and either untreated or exposed to 2mM of CoSO4 for 60 min. Changes in the transcriptome of yap1 mutant cells were then analyzed.
Project description:Growth assay in the presence of a toxic chemical that uses the barcoded collections of yeast gene deletions (haploid, diploid, DamP) to identify deletion strains that are hypersensitive to the drug.
Project description:Yeast is a powerful model system for studying the action of small molecule therapeutics. An important limitation has been low efficacy of many small molecules in yeast due to limited intracellular drug accumulation. We used the DNA binding domain of the pleiotropic drug resistance regulator Pdr1 fused in-frame to transcription repressors to repress Pdr1 regulated genes. Expression of these regulators conferred dominant enhancement of drug sensitivity and led to greatly diminished levels of Pdr1p regulated transcripts, including the yeast p-glycoprotein homologue Pdr5. Enhanced sensitivity was seen for a wide range of small molecules. Biochemical measurements demonstrated enhanced accumulation of rhodamine in yeast cells carrying the chimeras. These repressors of Pdr1p regulated transcripts can be introduced into large collections of strains such as the S. cerevisiae deletion set, and enhance the utility of yeast for studying drug action and for mechanism-based drug discovery. Experiment Overall Design: We determined the profiles of gene expression in yeast strains expressing dominant-negative Pdr1-fusion transcription factors. Yeast strains expressing different Pdr1-fusion repressors were analyzed by extracting total RNA and hybridization to Affymetrix GeneChip YG-s98 microarrays
