title: Quantification of genetically controlled cell death in budding yeast.
abstract: Yeast are the foremost genetic model system. With relative ease, entire chemical libraries can be screened for effects on essentially every gene in the yeast genome. Until recently, researchers focused only on whether yeast were killed by the conditions applied, irrespective of the mechanisms by which they died. In contrast, considerable effort has been devoted to understanding the mechanisms of mammalian cell death. However, most of the methodologies for detecting programmed apoptotic and necrotic death of mammalian cells have not been applicable to yeast. Therefore, we developed a cell death assay for baker's yeast Saccharomyces cerevisiae to identify genes involved in the mechanisms of yeast cell death. Small volumes of yeast suspensions are subjected to a precisely controlled heat ramp, allowing sufficient time for yeast cell factors to suppress or facilitate death, which can be quantified by high-throughput automated analyses. This assay produces remarkably reliable results that typically reflect results with other death stimuli. Here we describe the protocol and its caveats, which can be easily overcome.
authors.affiliation: W. Harry Feinstone Department of Molecular Microbiology and Immunology, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, MD, USA.
authors.forename: Xinchen
authors.initials: X
authors.lastname: Teng
authors.forename: J Marie
authors.initials: JM
authors.lastname: Hardwick
mesh_terms: D016923:Cell Death; D015169:Colony Count, Microbial; D003584:Cytological Techniques / Q000379:methods*; D006358:Hot Temperature; D050296:Microbial Viability; D012441:Saccharomyces cerevisiae / Q000166:cytology* / Q000235:genetics* / Q000254:growth & development; D013194:Staining and Labeling