Modeling Metazoan Apoptotic Pathways in Yeast.
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.
Department of Biochemistry and Genetics, La Trobe Institute for Molecular Science, La Trobe University, Bundoora, Melbourne, VIC, 3083, Australia.
David T
DT
Bloomer
Department of Biochemistry and Genetics, La Trobe Institute for Molecular Science, La Trobe University, Bundoora, Melbourne, VIC, 3083, Australia.
Tanja
T
Kitevska
Department of Biochemistry and Genetics, La Trobe Institute for Molecular Science, La Trobe University, Bundoora, Melbourne, VIC, 3083, Australia.
Ingo L
IL
Brand
Murdoch Children's Research Institute, Royal Children's Hospital, Parkville, VIC, 3052, Australia.|Australian Centre for Blood Diseases, Faculty of Medicine, Nursing and Health Science, Monash University, Melbourne, VIC, 3004, Australia.
Anissa M
AM
Jabbour
Department of Biochemistry and Genetics, La Trobe Institute for Molecular Science, La Trobe University, Bundoora, Melbourne, VIC, 3083, Australia.
Hang
H
Nguyen
Department of Biochemistry and Genetics, La Trobe Institute for Molecular Science, La Trobe University, Bundoora, Melbourne, VIC, 3083, Australia. c.hawkins@latrobe.edu.au.|Murdoch Children's Research Institute, Royal Children's Hospital, Parkville, VIC, 3052, Australia. c.hawkins@latrobe.edu.au.|Department of Biochemistry and Genetics, La Trobe Institute for Molecular Science, La Trobe University, Kingsbury Drive, Bundoora, Melbourne, VIC, 3083, Australia. c.hawkins@latrobe.edu.au.
Christine J
CJ
Hawkins
D000818:Animals; D017209:Apoptosis; D051017:Apoptosis Regulatory Proteins / Q000235:genetics / Q000378:metabolism; D020169:Caspases / Q000378:metabolism; D017930:Genes, Reporter; D007763:Lac Operon; D008928:Mitochondria / Q000378:metabolism; D008954:Models, Biological; D012441:Saccharomyces cerevisiae / Q000166:cytology* / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D015398:Signal Transduction; D014157:Transcription Factors / Q000378:metabolism
D051017:Apoptosis Regulatory Proteins; D014157:Transcription Factors; D020169:Caspases
Apoptosis; BH3-mimetic; Bax; Bcl-2; Caspase; IAP; Programmed cell death; Saccharomyces cerevisiae; Yeast