Genome-Wide Screen of Saccharomyces cerevisiae to Identify Genes Sensitive to Rapamycin and Involved in TOR Signaling Pathways
Samantha Parker, South Side High School, Rockville Centre; Bruce Futcher, Department of Molecular Genetics and Microbiology, Stony Brook University

The potent anti-inflammatory drug rapamycin has been shown to be an inhibitor of the Target Of Rapamycin (TOR) phosphatidylinositol 3-kinase. In the budding yeast Saccharomyces cerevisiae the two TOR homologues, TOR1 and TOR2, mediate a variety of cellular processes including growth and metabolism. As TOR is conserved in eukaryotes, knowledge of downstream TOR targets in yeast may elucidate the biological basis of TOR's effects on processes such as growth, metabolism, autophagy, and inflammation in other eukaryotes. Utilizing the yeast deletion set, ~4800 defined deletion strains were tested for rapamycin sensitivity. Strains were tested for degree of sensitivity and for rapamycin-specific sensitivity by using Hydroxyurea ( a G1-S inhibitor) at low concentrations. The majority of yeast strains were found to be relatively rapamycin insensitive at low concentrations (<7.5ng/ml); about ~2% of all strains displayed rapamycin-specific sensitivity. At high concentrations (>25ng/ml) ~1% of strains showed resistance. Funding was provided by the Simons Foundation.

 

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