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Exploration of essential gene functions via titratable promoter alleles |
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Sanie Mnaimneh*,1, Armaity P. Davierwala*,1, Jennifer Haynes*,2, Jason Moffat2, Wen-Tao Peng1, Wen Zhang1,2, Xueqi Yang1, Jeff Pootoolal1, Gordon Chua1, Andres Lopez1, Miles Trochesset1, Darcy Morse3, Nevan J. Krogan1,2, Shawna L. Hiley1, Zhijian Li1,2, Quaid Morris1, Jörg Grigull1, Nicholas Mitsakakis1, Christopher J. Roberts4, Jack F. Greenblatt1,2, Charles Boone1,2, Chris A. Kaiser3, Brenda J. Andrews2, and Timothy R. Hughes1,2§ |
*Indicates equal contribution
1Banting and Best Department of Medical Research,
University of Toronto, 112 College St., Toronto, ON M5G 1L6 Canada
2Department of Medical Genetics and Microbiology, University of
Toronto, 10 Kings College Circle, Toronto, ON Canada
3Department of Biology, Massachusetts Institute of Technology,
Cambridge, MA 02139, USA
4Rosetta Inpharmatics LLC, a wholly owned subsidiary of Merck &
Co., Inc., 401 Terry Ave. N., Seattle, WA 98109, USA.
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Abstract: Nearly 20% of yeast genes are required for viability, hindering genetic analysis with knockouts. We created promoter-shutoff strains for over two-thirds of all essential yeast genes and subjected them to morphological analysis, size profiling, drug sensitivity screening, and microarray expression profiling. We then used this compendium of data to ask which phenotypic features characterized different functional classes, and used these to infer potential functions for uncharacterized genes. We identified genes involved in ribosome biogenesis (HAS1, URB1 and URB2), protein secretion (SEC39), mitochondrial import (MIM1), and tRNA charging (GSN1). In addition, apparent negative-feedback transcriptional regulation of both ribosome biogenesis and the proteasome was observed. We furthermore show that these strains are compatible with automated genetic analysis. This study underscores the importance of analyzing mutant phenotypes, and provides a resource to complement the yeast knockout collection. |
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