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Activation of the mitochondrial death pathway is commonly mediated by a preferential engagement of Bak

Oncogene volume 27, pages 1387–1396 (2008)Cite this article

Abstract

Among the members of the Bcl-2 family, the multidomain proteins Bax and Bak are crucial for the activation of mitochondria. However, it is still unclear whether they act in a unique and distinct manner or whether they exhibit redundant functions. To systematically investigate their activation on a single-cell level, we established MCF-7 cell lines stably expressing GFP-fusion variants of these proteins. We found that MCF-7/GFP-Bak cells showed an increased sensitivity to apoptosis induction by staurosporine, actinomycin D, TRAIL and overexpression of Puma compared to GFP-Bax-expressing cells. Independently of the death stimulus used, oligomerization of endogenous and exogenous Bak was mostly detected prior to an activation of Bax, whereas cells displaying oligomerized Bax in the absence of Bak clusters were not observed. In addition, activation of Bax but not Bak was attenuated by a caspase inhibitor. Consistent with this, caspase-3-deficient MCF-7 cells displayed a significantly reduced activation of endogenous Bax than caspase-3-proficient MCF-7 cells. Thus, our data strongly suggest that diverse apoptotic stimuli preferentially engage the Bak pathway, whereas the triggering of Bax occurs, at least partially, downstream of mitochondrial caspase activation, most likely constituting a positive feedback loop for the amplification of the death signal.

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Acknowledgements

We thank Andrea Meyer for expert technical assistance and Klaus Meyer for cell sorting. This work was supported by grants from the Deutsche Forschungsgemeinschaft (SFB 728) to RUJ and the Forschungskommission of the University of Düsseldorf to FE.

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Authors and Affiliations

  1. Institute of Molecular Medicine, University of Düsseldorf, Düsseldorf, Germany

    D Neise, V Graupner, K Schulze-Osthoff, R U Jänicke & F Essmann

  2. Department of Hematology, Oncology and Tumor Immunology, University Medical Center Charité, Berlin, Germany

    B F Gillissen & P T Daniel

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  1. D Neise
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Correspondence to F Essmann.

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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc).

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Neise, D., Graupner, V., Gillissen, B. et al. Activation of the mitochondrial death pathway is commonly mediated by a preferential engagement of Bak. Oncogene 27, 1387–1396 (2008). https://doi.org/10.1038/sj.onc.1210773

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