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Quantification of Genetically Controlled Cell Death in Budding Yeast

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Necrosis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1004))

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.

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Acknowledgment

This work was supported by NIH grant GM077875 (J.M.H.).

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Authors and Affiliations

  1. W. Harry Feinstone Department of Molecular Microbiology and Immunology, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, MD, USA

    Xinchen Teng & J. Marie Hardwick

Authors
  1. Xinchen Teng
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  2. J. Marie Hardwick
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Editor information

Editors and Affiliations

  1. Department of Biology, Boston University, Boston, MA, USA

    Kimberly McCall

  2. Brookline, MA, USA

    Charles Klein

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Teng, X., Hardwick, J.M. (2013). Quantification of Genetically Controlled Cell Death in Budding Yeast. In: McCall, K., Klein, C. (eds) Necrosis. Methods in Molecular Biology, vol 1004. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-383-1_12

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  • DOI: https://doi.org/10.1007/978-1-62703-383-1_12

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-382-4

  • Online ISBN: 978-1-62703-383-1

  • eBook Packages: Springer Protocols

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