title: High-throughput phenotypic profiling of gene-environment interactions by quantitative growth curve analysis in Saccharomyces cerevisiae.
abstract: Cell-based assays are widely used in high-throughput screening to determine the effects of toxicants and drugs on their biological targets. To enable a functional genomics modeling of gene-environment interactions, quantitative assays are required both for gene expression and for the phenotypic responses to environmental challenge. To address this need, we describe an automated high-throughput methodology that provides phenotypic profiling of the cellular responses to environmental stress in Saccharomyces cerevisiae. Standardized assay conditions enable the use of a single metric value to quantify yeast microculture growth curves. This assay format allows precise control of both genetic and environmental determinants of the cellular responses to oxidative stress, a common mechanism of environmental insult. These yeast-cell-based assays are validated with hydrogen peroxide, a simple direct-acting oxidant. Phenotypic profiling of the oxidative stress response of a yap1 mutant strain demonstrates the mechanistic analysis of genetic susceptibility to oxidative stress. As a proof of concept for analysis of more complex gene-environment interactions, we describe a combinatorial assay design for phenotypic profiling of the cellular responses to tert-butyl hydroperoxide, a complex oxidant that is actively metabolized by its target cells. Thus, the yeast microculture assay format supports comprehensive applications in toxicogenomics.
authors.affiliation: Center for Molecular Medicine and Genetics, Wayne State University, Detroit, MI 48202, USA.
authors.forename: Andrew
authors.initials: A
authors.lastname: Weiss
authors.forename: James
authors.initials: J
authors.lastname: Delproposto
authors.forename: Craig N
authors.initials: CN
authors.lastname: Giroux
mesh_terms: D004777:Environment; D006861:Hydrogen Peroxide / Q000378:metabolism / Q000494:pharmacology; D008904:Miniaturization; D018384:Oxidative Stress; D010597:Pharmacogenetics / Q000379:methods*; D010641:Phenotype; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics; D017384:Sequence Deletion; D014157:Transcription Factors / Q000235:genetics; D020122:tert-Butylhydroperoxide / Q000378:metabolism / Q000494:pharmacology
chemical_list: D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; C086677:YAP1 protein, S cerevisiae; D020122:tert-Butylhydroperoxide; D006861:Hydrogen Peroxide