High-throughput genetic interaction mapping in the fission yeast Schizosaccharomyces pombe.
Epistasis analysis, which reports on the extent to which the function of one gene depends on the presence of a second, is a powerful tool for studying the functional organization of the cell. Systematic genome-wide studies of epistasis, however, have been limited, with the majority of data being collected in the budding yeast, Saccharomyces cerevisiae. Here we present two 'pombe epistasis mapper' strategies, PEM-1 and PEM-2, which allow for high-throughput double mutant generation in the fission yeast, S. pombe. These approaches take advantage of a previously undescribed, recessive, cycloheximide-resistance mutation. Both systems can be used for genome-wide screens or for the generation of high-density, quantitative epistatic miniarray profiles (E-MAPs). Since S. cerevisiae and S. pombe are evolutionary distant, this methodology will provide insight into conserved biological pathways that are present in S. pombe, but not S. cerevisiae, and will enable a comprehensive analysis of the conservation of genetic interaction networks.
Roguev
Assen
A
Department of Cellular and Molecular Pharmacology, University of California, San Francisco, 1700 4th Street San Francisco, California 94158, USA.
Wiren
Marianna
M
Weissman
Jonathan S
JS
Krogan
Nevan J
NJ
D004843:Epistasis, Genetic; D005804:Genes, Lethal / Q000235:genetics; D016681:Genome, Fungal / Q000235:genetics*; D023281:Genomics / Q000379:methods*; D009154:Mutation; D012568:Schizosaccharomyces / Q000235:genetics*; D014170:Transformation, Genetic