Mito-priming as a method to engineer Bcl-2 addiction

Most apoptotic stimuli require mitochondrial outer membrane permeabilization (MOMP) in order to execute cell death. As such, MOMP is subject to tight control by Bcl-2 family proteins. We have developed a powerful new technique to investigate Bcl-2-mediated regulation of MOMP. This method, called mit...

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Bibliographic Details
Published in:Nature communications 2016-02, Vol.7 (1), p.10538-10538, Article 10538
Main Authors: Lopez, Jonathan, Bessou, Margaux, Riley, Joel S., Giampazolias, Evangelos, Todt, Franziska, Rochegüe, Tony, Oberst, Andrew, Green, Douglas R., Edlich, Frank, Ichim, Gabriel, Tait, Stephen W. G.
Format: Article
Language:English
Subjects:
13/2
14/19
631/1647/1407
631/80/82/23
631/80/86
Animals
Apoptosis - physiology
Apoptosis Regulatory Proteins - genetics
Apoptosis Regulatory Proteins - metabolism
Cell Line
CRISPR-Cas Systems
Gene Expression Regulation
Genetic Engineering
Humanities and Social Sciences
Humans
Mice
Mitochondrial Membranes - physiology
multidisciplinary
Peptide Fragments - metabolism
Proto-Oncogene Proteins - metabolism
Proto-Oncogene Proteins c-bcl-2 - genetics
Proto-Oncogene Proteins c-bcl-2 - metabolism
Science
Science (multidisciplinary)
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Summary:Most apoptotic stimuli require mitochondrial outer membrane permeabilization (MOMP) in order to execute cell death. As such, MOMP is subject to tight control by Bcl-2 family proteins. We have developed a powerful new technique to investigate Bcl-2-mediated regulation of MOMP. This method, called mito-priming, uses co-expression of pro- and anti-apoptotic Bcl-2 proteins to engineer Bcl-2 addiction. On addition of Bcl-2 targeting BH3 mimetics, mito-primed cells undergo apoptosis in a rapid and synchronous manner. Using this method we have comprehensively surveyed the efficacy of BH3 mimetic compounds, identifying potent and specific MCL-1 inhibitors. Furthermore, by combining different pro- and anti-apoptotic Bcl-2 pairings together with CRISPR/Cas9-based genome editing, we find that tBID and PUMA can preferentially kill in a BAK-dependent manner. In summary, mito-priming represents a facile and robust means to trigger mitochondrial apoptosis. Apoptosis often requires mitochondrial outer membrane permeabilization, a process targeted by Bcl-2-binding BH3 mimetics. Here the authors describe and apply 'mito-priming', a method that allows triggering mitochondrial apoptosis in a synchronous manner, facilitating the investigation of mitochondrial apoptosis and its regulation by Bcl-2 proteins.
ISSN:2041-1723
2041-1723
DOI:10.1038/ncomms10538