Quantification of genetically controlled cell death in budding yeast

Xinchen Teng, J. Marie Hardwick

Research output: Chapter in Book/Report/Conference proceedingChapter

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.

Original languageEnglish (US)
Title of host publicationNecrosis
Subtitle of host publicationMethods and Protocols
PublisherHumana Press Inc.
Pages161-170
Number of pages10
ISBN (Print)9781627033824
DOIs
StatePublished - 2013

Publication series

NameMethods in Molecular Biology
Volume1004
ISSN (Print)1064-3745

Keywords

  • Apoptosis
  • Cell death
  • Heat ramp
  • Necrosis
  • Yeast

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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