Abstract
Necroptosis is a form of programmed cell death that occurs in the absence of caspase activation and depends on the activity of the receptor-interacting protein kinases. Inactivation of these kinases by caspase-mediated cleavage has been shown to be essential for successful embryonic development, survival and activation of certain cell types. The initiator of extrinsic apoptosis, caspase-8, which has a pro-death as well as a pro-life function, has been assigned this role. In the present study we demonstrate that caspase-6, an executioner caspase, performs this role during apoptosis induced through the intrinsic pathway. In addition, we demonstrate that in the absence of caspase activity, intrinsic triggers of apoptosis induce the receptor-interacting-kinase-1-dependent production of pro-inflammatory cytokines. We show that ubiquitously expressed caspase-6 has a supporting role in apoptosis by cleaving this kinase, thus preventing production of inflammatory cytokines as well as inhibiting the necroptotic pathway. These findings shed new light on the regulation of necroptosis as well as cell death in an inflammatory environment wherein cells receive both intrinsic and extrinsic death signals.
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Abbreviations
- AV:
-
Annexin V
- bEVD-AOMK:
-
biotinyl-GluValAsp-acyloxymethyl ketone
- cFLIP:
-
cellular FLICE-like inhibitory protein
- DSBs:
-
double strand breaks
- FKBP:
-
FK506 binding protein
- Nec1:
-
necrostatin-1
- RIPK:
-
receptor-interacting protein kinase
- SytR:
-
Sytox Red
- zVAD:
-
carbobenzoxy-ValAlaAsp-fluoromethyl ketone
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Acknowledgements
We thank Matt Bogyo, Stanford, for the gift of bEVD-AOMK. We gratefully acknowledge Scott Snipas, Insuk Andersen and Xiaofan Qiu for technical assistance and the other members of the Salvesen lab, as well as Stefan Riedl and Peter Mace, for insightful discussions. This work was supported by NIH R01-GM099040. BJvR is supported by a Rubicon fellowship from the Netherlands Organization for Scientific Research as well as a fellowship from the Barth Syndrome Foundation. DEE is supported by a Canadian Institutes of Health Research (CIHR) fellowship, the work was further supported by grants from CIHR (MOP-84438) and the Cure Huntington’s Disease Initiative (Treat-HD) to MRH.
Author Contributions
BJvR performed and BJvR and GSS designed experiments, analyzed data and wrote the manuscript. DEE performed and DEE and MRH designed the experiments and analyzed the data shown in Figure 4.
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van Raam, B., Ehrnhoefer, D., Hayden, M. et al. Intrinsic cleavage of receptor-interacting protein kinase-1 by caspase-6. Cell Death Differ 20, 86–96 (2013). https://doi.org/10.1038/cdd.2012.98
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DOI: https://doi.org/10.1038/cdd.2012.98
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