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.
Mnaimneh
Sanie
S
Banting and Best Department of Medical Research, University of Toronto, 112 College Street, Toronto, ON M5G 1L6, Canada.
Davierwala
Armaity P
AP
Haynes
Jennifer
J
Moffat
Jason
J
Peng
Wen-Tao
WT
Zhang
Wen
W
Yang
Xueqi
X
Pootoolal
Jeff
J
Chua
Gordon
G
Lopez
Andres
A
Trochesset
Miles
M
Morse
Darcy
D
Krogan
Nevan J
NJ
Hiley
Shawna L
SL
Li
Zhijian
Z
Morris
Quaid
Q
Grigull
Jörg
J
Mitsakakis
Nicholas
N
Roberts
Christopher J
CJ
Greenblatt
Jack F
JF
Boone
Charles
C
Kaiser
Chris A
CA
Andrews
Brenda J
BJ
Hughes
Timothy R
TR
D000483:Alleles; D025461:Feedback, Physiological; D017353:Gene Deletion; D020869:Gene Expression Profiling; D015966:Gene Expression Regulation, Fungal; D020043:Genes, Essential; D005800:Genes, Fungal; D008928:Mitochondria / Q000378:metabolism; D008957:Models, Genetic; D020411:Oligonucleotide Array Sequence Analysis; D004364:Pharmaceutical Preparations / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011499:Protein Processing, Post-Translational; D012343:RNA, Transfer / Q000378:metabolism; D012269:Ribosomal Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic
D004364:Pharmaceutical Preparations; D012269:Ribosomal Proteins; D029701:Saccharomyces cerevisiae Proteins; D012343:RNA, Transfer