Modeling metazoan apoptotic pathways in yeast

David T Bloomer, Tanja Kitevska, Ingo L Brand, Anissa M Jabbour, Hang Nguyen, Christine J Hawkins

Research output: Chapter in Book/Report/Conference proceedingChapter (Book)Otherpeer-review

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.
Original languageEnglish
Title of host publicationProgrammed Cell Death
Subtitle of host publicationMethods and Protocols
EditorsHamsa Puthalakath, Christine J. Hawkins
Place of PublicationNew York NY USA
PublisherHumana Press
Pages161 - 183
Number of pages23
ISBN (Print)978-1-4939-3579-6
DOIs
Publication statusPublished - 2016

Publication series

NameMethods in Molecular Biology
PublisherSpringer
Volume1419
ISSN (Print)1064-3745

Cite this

Bloomer, D. T., Kitevska, T., Brand, I. L., Jabbour, A. M., Nguyen, H., & Hawkins, C. J. (2016). Modeling metazoan apoptotic pathways in yeast. In H. Puthalakath, & C. J. Hawkins (Eds.), Programmed Cell Death: Methods and Protocols (pp. 161 - 183). (Methods in Molecular Biology; Vol. 1419). New York NY USA: Humana Press. https://doi.org/10.1007/978-1-4939-3581-9_13
Bloomer, David T ; Kitevska, Tanja ; Brand, Ingo L ; Jabbour, Anissa M ; Nguyen, Hang ; Hawkins, Christine J. / Modeling metazoan apoptotic pathways in yeast. Programmed Cell Death: Methods and Protocols. editor / Hamsa Puthalakath ; Christine J. Hawkins. New York NY USA : Humana Press, 2016. pp. 161 - 183 (Methods in Molecular Biology).
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Bloomer, DT, Kitevska, T, Brand, IL, Jabbour, AM, Nguyen, H & Hawkins, CJ 2016, Modeling metazoan apoptotic pathways in yeast. in H Puthalakath & CJ Hawkins (eds), Programmed Cell Death: Methods and Protocols. Methods in Molecular Biology, vol. 1419, Humana Press, New York NY USA, pp. 161 - 183. https://doi.org/10.1007/978-1-4939-3581-9_13

Modeling metazoan apoptotic pathways in yeast. / Bloomer, David T; Kitevska, Tanja; Brand, Ingo L; Jabbour, Anissa M; Nguyen, Hang; Hawkins, Christine J.

Programmed Cell Death: Methods and Protocols. ed. / Hamsa Puthalakath; Christine J. Hawkins. New York NY USA : Humana Press, 2016. p. 161 - 183 (Methods in Molecular Biology; Vol. 1419).

Research output: Chapter in Book/Report/Conference proceedingChapter (Book)Otherpeer-review

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AB - 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.

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Bloomer DT, Kitevska T, Brand IL, Jabbour AM, Nguyen H, Hawkins CJ. Modeling metazoan apoptotic pathways in yeast. In Puthalakath H, Hawkins CJ, editors, Programmed Cell Death: Methods and Protocols. New York NY USA: Humana Press. 2016. p. 161 - 183. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-3581-9_13