Exploration of essential gene functions via titratable promoter alleles

Sanie Mnaimneh, Armaity P. Davierwala, Jennifer Haynes, Jason Moffat, Wen Tao Peng, Wen Zhang, Xueqi Yang, Jeff Pootoolal, Gordon Chua, Andres Lopez, Miles Trochesset, Darcy Morse, Nevan J. Krogan, Shawna L. Hiley, Zhijian Li, Quaid Morris, Jörg Grigull, Nicholas Mitsakakis, Christopher J. Roberts, Jack F. GreenblattCharles Boone, Chris A. Kaiser, Brenda J. Andrews, Timothy R. Hughes

Research output: Contribution to journalArticlepeer-review

489 Scopus citations

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.

Original languageEnglish
Pages (from-to)31-44
Number of pages14
JournalCell
Volume118
Issue number1
DOIs
StatePublished - 9 Jul 2004
Externally publishedYes

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