Exploration of essential gene functions via titratable promoter alleles.
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.
Pubmed ID: 15242642 RIS Download
Alleles | Feedback, Physiological | Gene Deletion | Gene Expression Profiling | Gene Expression Regulation, Fungal | Genes, Essential | Genes, Fungal | Mitochondria | Models, Genetic | Oligonucleotide Array Sequence Analysis | Pharmaceutical Preparations | Promoter Regions, Genetic | Protein Processing, Post-Translational | RNA, Transfer | Ribosomal Proteins | Saccharomyces cerevisiae | Saccharomyces cerevisiae Proteins | Transcription, Genetic