A comprehensive strategy enabling high-resolution functional analysis of the yeast genome.
Functional genomic studies in Saccharomyces cerevisiae have contributed enormously to our understanding of cellular processes. Their full potential, however, has been hampered by the limited availability of reagents to systematically study essential genes and the inability to quantify the small effects of most gene deletions on growth. Here we describe the construction of a library of hypomorphic alleles of essential genes and a high-throughput growth competition assay to measure fitness with unprecedented sensitivity. These tools dramatically increase the breadth and precision with which quantitative genetic analysis can be performed in yeast. We illustrate the value of these approaches by using genetic interactions to reveal new relationships between chromatin-modifying factors and to create a functional map of the proteasome. Finally, by measuring the fitness of strains in the yeast deletion library, we addressed an enigma regarding the apparent prevalence of gene dispensability and found that most genes do contribute to growth.
Breslow
David K
DK
Department of Cellular and Molecular Pharmacology, University of California, San Francisco, 1700 4th Street, San Francisco, California 94158, USA.
Cameron
Dale M
DM
Collins
Sean R
SR
Schuldiner
Maya
M
Stewart-Ornstein
Jacob
J
Newman
Heather W
HW
Braun
Sigurd
S
Madhani
Hiten D
HD
Krogan
Nevan J
NJ
Weissman
Jonathan S
JS
D000483:Alleles; D030561:Databases, Nucleic Acid; D005434:Flow Cytometry; D017353:Gene Deletion; D005821:Genetic Techniques; D016681:Genome, Fungal / Q000235:genetics*; D046988:Proteasome Endopeptidase Complex / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D012680:Sensitivity and Specificity
D029701:Saccharomyces cerevisiae Proteins; D046988:Proteasome Endopeptidase Complex