Modeling Metazoan Apoptotic Pathways in Yeast

Methods Mol Biol. 2016:1419:161-83. doi: 10.1007/978-1-4939-3581-9_13.

Abstract

This chapter describes techniques for characterizing metazoan apoptotic pathways using Saccharomyces cerevisiae. Active forms of the major apoptotic effectors-caspases, Bax and Bak-are all lethal to yeast. Using this lethality as a readout of caspase/Bax/Bak activity, proteins and small molecules that directly or indirectly regulate the activity of these effectors can be investigated in yeast, and apoptotic inhibitors can be identified using functional yeast-based screens. Caspase activity can also be monitored in yeast by cleavage-dependent liberation of a transcription factor from the plasma membrane, enabling it to activate the lacZ reporter gene. This system can be used to define the sequences that can be efficiently cleaved by particular caspases.

Keywords: Apoptosis; BH3-mimetic; Bax; Bcl-2; Caspase; IAP; Programmed cell death; Saccharomyces cerevisiae; Yeast.

MeSH terms

  • Animals
  • Apoptosis Regulatory Proteins / genetics
  • Apoptosis Regulatory Proteins / metabolism
  • Apoptosis*
  • Caspases / metabolism
  • Genes, Reporter
  • Lac Operon
  • Mitochondria / metabolism
  • Models, Biological*
  • Saccharomyces cerevisiae / cytology*
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / growth & development
  • Saccharomyces cerevisiae / metabolism*
  • Signal Transduction*
  • Transcription Factors / metabolism

Substances

  • Apoptosis Regulatory Proteins
  • Transcription Factors
  • Caspases