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PMID: 18622397 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

A comprehensive strategy enabling high-resolution functional analysis of the yeast genome.

Nature methods ·Vol. 5 ·No. 8 ·2008-08-00 ·Pages 711-8

Breslow DK, Cameron DM, Collins SR, Schuldiner M, Stewart-Ornstein J, Newman HW, Braun S, Madhani HD, Krogan NJ, Weissman JS

Abstract

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.

MeSH Terms
Alleles Databases, Nucleic Acid Flow Cytometry Gene Deletion Genetic Techniques Genome, Fungal/genetics Proteasome Endopeptidase Complex/genetics,metabolism Saccharomyces cerevisiae/genetics,metabolism Saccharomyces cerevisiae Proteins/analysis,genetics,metabolism Sensitivity and Specificity
Chemicals
Saccharomyces cerevisiae Proteins Proteasome Endopeptidase Complex
Authors & Affiliations
10 authors, click to expand affiliations / ORCID
Breslow David K
Department of Cellular and Molecular Pharmacology, University of California, San Francisco, 1700 4th Street, San Francisco, California 94158, USA.
Cameron Dale M
Collins Sean R
Schuldiner Maya
Stewart-Ornstein Jacob
Newman Heather W
Braun Sigurd
Madhani Hiten D
Krogan Nevan J
Weissman Jonathan S
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Article Info
Journal
Nature methods
Abbr.
Nat Methods
ISSN
1548-7105
DOI
10.1038/nmeth.1234
Published
2008-08-00
Epub
2008-00-11
Pages
711-8
Language
English
Region
United States
NLM ID
101215604
PMCID
PMC2756093
Subset
IM
Grants
Howard Hughes Medical Institute · United States
External Links
PubMed source DOI: 10.1038/nmeth.1234 PMC Full Text View journal details
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