TY - JOUR
T1 - Microarray-based method for monitoring yeast overexpression strains reveals small-molecule targets in TOR pathway
AU - Butcher, Rebecca A.
AU - Bhullar, Bhupinder S.
AU - Perlstein, Ethan O.
AU - Marsischky, Gerald
AU - LaBaer, Joshua
AU - Schreiber, Stuart L.
N1 - Funding Information:
The authors thank M. Hall for the gift of the fpr1-8 strain, C.L. Liu for advice regarding the in vitro transcription amplification strategy and for help with data analysis, and A. Shamji for a careful reading of this manuscript. This work was supported by GM38627 (awarded to S.L.S.). The construction of the collection of yeast overexpression plasmids was supported by National Human Genome Research Institute R01-HG002923 (awarded to J.L.). S.L.S. is an Investigator at the Howard Hughes Medical Institute. R.A.B. was supported by a graduate fellowship from the National Science Foundation.
PY - 2006/2
Y1 - 2006/2
N2 - Identification of the cellular targets of small-molecule hits in phenotypic screens is a central challenge in the development of small molecules as biological tools and potential therapeutics. To facilitate the process of small-molecule target identification, we developed a global, microarray-based method for monitoring the growth of pools of yeast strains, each overexpressing a different protein, in the presence of small molecules. Specifically, the growth of Saccharomyces cerevisiae strains harboring ∼3,900 different overexpression plasmids was monitored in the presence of rapamycin, which inhibits the target of rapamycin (TOR) proteins. TOR was successfully identified as a candidate rapamycin target, and many additional gene products were implicated in the TOR signaling pathway. We also characterized the mechanism of LY-83583, a small-molecule suppressor of rapamycin-induced growth inhibition. These data enabled functional links to be drawn between groups of genes implicated in the TOR pathway, identified several candidate targets for LY-83583, and suggested a role for mitochondrial respiration in mediating rapamycin sensitivity.
AB - Identification of the cellular targets of small-molecule hits in phenotypic screens is a central challenge in the development of small molecules as biological tools and potential therapeutics. To facilitate the process of small-molecule target identification, we developed a global, microarray-based method for monitoring the growth of pools of yeast strains, each overexpressing a different protein, in the presence of small molecules. Specifically, the growth of Saccharomyces cerevisiae strains harboring ∼3,900 different overexpression plasmids was monitored in the presence of rapamycin, which inhibits the target of rapamycin (TOR) proteins. TOR was successfully identified as a candidate rapamycin target, and many additional gene products were implicated in the TOR signaling pathway. We also characterized the mechanism of LY-83583, a small-molecule suppressor of rapamycin-induced growth inhibition. These data enabled functional links to be drawn between groups of genes implicated in the TOR pathway, identified several candidate targets for LY-83583, and suggested a role for mitochondrial respiration in mediating rapamycin sensitivity.
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U2 - 10.1038/nchembio762
DO - 10.1038/nchembio762
M3 - Article
C2 - 16415861
AN - SCOPUS:33644899143
SN - 1552-4450
VL - 2
SP - 103
EP - 109
JO - Nature Chemical Biology
JF - Nature Chemical Biology
IS - 2
ER -