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Necrosis-like death can engage multiple pro-apoptotic Bcl-2 protein family members

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Abstract

Necroptosis is a physiologically relevant mode of cell death with some well-described initiating events, but largely unknown executioners. Here we investigated necrostatin-1 (Nec-1) sensitive death elicited by different necroptosis stimuli in L929 mouse fibrosarcoma cells, mouse embryonic fibroblasts (MEF) and bone marrow-derived macrophages. We found that TNFα- or zVAD-induced necroptosis occurs independently of the recently implicated executioners Bmf or PARP-2, but can involve the Bcl-2 family proteins Bid and Bak. Furthermore, this type of necroptosis is associated with mitochondrial cytochrome c release and partly sensitive to cyclosporine A inhibition, suggesting a cross talk with the mitochondrial permeability transition pore. Necroptosis triggered by cadmium (Cd) exposure caused fully Nec-1-sensitive and caspase-independent death in L929 cells that was associated with autocrine TNFα-mediated feed-forward signalling. In MEF Cd-exposure elicited a mixed mode of cell death that was to some extent Nec-1-sensitive but also displayed features of apoptosis. It was partly dependent on Bmf and Bax/Bak, proteins typically considered to act pro-apoptotic, but ultimately insensitive to caspase inhibition. Overall, our study indicates that inducers of “extrinsic” and “intrinsic” necroptosis can both trigger TNF-receptor signalling. Further, necroptosis may depend on mitochondrial changes engaging proteins considered critical for MOMP during apoptosis that ultimately contribute to caspase-independent necrotic cell death.

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Acknowledgments

We thank J Silke, M Kelliher, A Strasser, J Adams, R Flavell and Y Jelamos for providing MEF, mice and/or reagents. We acknowledge skilful technical assistance by Katharina Heinz and Eva Albertini. This study was supported by grants from the Austrian Science Fund (FWF) # Y212-B13 START (AV), the Tiroler Krebshilfe (CM, GK) and the Tyrolean Science Fund (CM). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Division of Developmental Immunology, Biocenter, Medical University Innsbruck, 6020, Innsbruck, Austria

    Denise Tischner, Claudia Manzl, Claudia Soratroi, Andreas Villunger & Gerhard Krumschnabel

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  1. Denise Tischner
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  2. Claudia Manzl
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  3. Claudia Soratroi
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  4. Andreas Villunger
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  5. Gerhard Krumschnabel
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Correspondence to Gerhard Krumschnabel.

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Denise Tischner and Claudia Manzl contributed equally to this work.

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Tischner, D., Manzl, C., Soratroi, C. et al. Necrosis-like death can engage multiple pro-apoptotic Bcl-2 protein family members. Apoptosis 17, 1197–1209 (2012). https://doi.org/10.1007/s10495-012-0756-8

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