title: Rapid identification of chemical genetic interactions in Saccharomyces cerevisiae.
abstract: Determining the mode of action of bioactive chemicals is of interest to a broad range of academic, pharmaceutical, and industrial scientists. Saccharomyces cerevisiae, or budding yeast, is a model eukaryote for which a complete collection of ~6,000 gene deletion mutants and hypomorphic essential gene mutants are commercially available. These collections of mutants can be used to systematically detect chemical-gene interactions, i.e. genes necessary to tolerate a chemical. This information, in turn, reports on the likely mode of action of the compound. Here we describe a protocol for the rapid identification of chemical-genetic interactions in budding yeast. We demonstrate the method using the chemotherapeutic agent 5-fluorouracil (5-FU), which has a well-defined mechanism of action. Our results show that the nuclear TRAMP RNA exosome and DNA repair enzymes are needed for proliferation in the presence of 5-FU, which is consistent with previous microarray based bar-coding chemical genetic approaches and the knowledge that 5-FU adversely affects both RNA and DNA metabolism. The required validation protocols of these high-throughput screens are also described.
authors: Department of Biochemistry and Microbiology, University of Victoria.
authors: David
authors: D
authors: Dilworth
authors: Department of Biochemistry and Microbiology, University of Victoria; cjn@uvic.ca.
authors: Christopher J
authors: CJ
authors: Nelson
mesh_terms: D000964:Antimetabolites, Antineoplastic / Q000494:pharmacology*; D045643:DNA Repair Enzymes / Q000378:metabolism; D004271:DNA, Fungal / Q000235:genetics / Q000378:metabolism; D005472:Fluorouracil / Q000494:pharmacology*; D059647:Gene-Environment Interaction; D012331:RNA, Fungal / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000187:drug effects* / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D000964:Antimetabolites, Antineoplastic; D004271:DNA, Fungal; D012331:RNA, Fungal; D029701:Saccharomyces cerevisiae Proteins; D045643:DNA Repair Enzymes; D005472:Fluorouracil