Project description:Transcriptional adaptation of yeast (BY4743 homozygous deletion mutant of HO) cells to long-term sustained exposure to rapamycin is investigated against the transcriptional profile of untreated cells. Cells were inoculated into 2nM rapamycin containing medium and were grown until mid-exponential phase. Both treated and untreated cells were grown in fermenters where the pH was controlled at 5.5 (pH1) or monitored (pH0), and dissolved oxygen saturation was controlled at >90% (air1) or monitored (air0) in a 2 x 2 factorial design. All experiments were conducted in duplicates (R1 and R2).
Project description:The target of rapamycin (TOR) plays a central role in eukaryotic cell growth control. With prevalent hyper-activation of the mTOR pathway in human cancers, novel strategies to enhance TOR pathway inhibition are highly desirable. We used a yeast-based high-throughput chemical genetic screen to identify small-molecule enhancers of rapamycin (SMERs) and used whole genome expression analysis to identify their mechanisms of action. We treated yeast individually with SMERs 1-5, rapamycin, or DMSO (solvent control) for 30 minutes prior to RNA extraction and hybridization on Affymetrix microarrays. Expression profiles of SMER-treated samples were compared to that of DMSO (solvent control) and rapamycin-treated samples to identify gene expression signatures unique to SMER-treated samples.
Project description:The target of rapamycin (TOR) plays a central role in eukaryotic cell growth control. With prevalent hyper-activation of the mTOR pathway in human cancers, novel strategies to enhance TOR pathway inhibition are highly desirable. We used a yeast-based high-throughput chemical genetic screen to identify small-molecule enhancers of rapamycin (SMERs) and used whole genome expression analysis to identify their mechanisms of action.
