Abstract
Three members of the IAP family (X-linked inhibitor of apoptosis (XIAP), cellular inhibitor of apoptosis proteins-1/-2 (cIAP1 and cIAP2)) are potent suppressors of apoptosis. Recent studies have shown that cIAP1 and cIAP2, unlike XIAP, are not direct caspase inhibitors, but block apoptosis by functioning as E3 ligases for effector caspases and receptor-interacting protein 1 (RIP1). cIAP-mediated polyubiquitination of RIP1 allows it to bind to the pro-survival kinase transforming growth factor-β-activated kinase 1 (TAK1) which prevents it from activating caspase-8-dependent death, a process reverted by the de-ubiquitinase CYLD. RIP1 is also a regulator of necrosis, a caspase-independent type of cell death. Here, we show that cells depleted of the IAPs by treatment with the IAP antagonist BV6 are greatly sensitized to tumor necrosis factor (TNF)-induced necrosis, but not to necrotic death induced by anti-Fas, poly(I:C) oxidative stress. Specific targeting of the IAPs by RNAi revealed that repression of cIAP1 is responsible for the sensitization. Similarly, lowering TAK1 levels or inhibiting its kinase activity sensitized cells to TNF-induced necrosis, whereas repressing CYLD had the opposite effect. We show that this sensitization to death is accompanied by enhanced RIP1 kinase activity, increased recruitment of RIP1 to Fas-associated via death domain and RIP3 (which allows necrosome formation), and elevated RIP1 kinase-dependent accumulation of reactive oxygen species (ROS). In conclusion, our data indicate that cIAP1 and TAK1 protect cells from TNF-induced necrosis by preventing RIP1/RIP3-dependent ROS production.
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Abbreviations
- 5Z-7:
-
5Z-7-oxozeaenol
- BHA:
-
butylated hydroxyanisole
- cIAP1/2:
-
cellular inhibitor of apoptosis proteins-1/-2
- CYLD:
-
cylindromatosis
- DHR123:
-
dihydrorhodamine 123
- ERK:
-
extracellular signal-regulated kinase
- FADD:
-
Fas-associated via death domain
- H2O2:
-
hydrogen peroxide
- IFNβ:
-
interferon-β
- MEK1/2:
-
mitogen-activated protein kinase kinase 1/-2
- NDUFB8:
-
NADH dehydrogenase (ubiquinone) 1 beta subcomplex 8
- Nec-1:
-
necrostatin-1
- Nox1:
-
NADPH oxidase 1
- RFK:
-
riboflavin kinase
- RIP:
-
receptor-interacting protein
- ROS:
-
reactive oxygen species
- TAK1:
-
transforming growth factor-β-activated kinase 1
- TLR3:
-
toll-like receptor 3
- TNF:
-
tumor necrosis factor
- TNFR1:
-
tumor necrosis factor receptor 1
- XIAP:
-
X-linked inhibitor of apoptosis
- zVAD-fmk:
-
benzyloxycarbonyl-Val-Ala-Asp(OMe)-fluoromethylketone
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Acknowledgements
We thank Professor Wim Declercq and Dr. Saskia Lippens for critical feedback and discussion and Dr. A Bredan for editing. We are grateful to Dr. RG Korneluk for sending the RIAP1 antibody. TV and MB received a postdoctoral fellowship from the FWO, PB is paid by VIB, and NV obtained a predoctoral fellowship from the BOF, Ghent University. BL was a master student working in the labs of both Professor Simone Fulda and Professor Peter Vandenabeele. Research in the Vandenabeele group is supported by VIB, Ghent University, Research Foundation Flanders (FWO-Vlaanderen) (3G.0218.06 and G.0226.09), Federal Research Program IAP 6/18, European Research Program FP6 ApopTrain (MRTN-CT-035624) and FP7 Apo-Sys 200767, and the GROUP-ID consortium of the UGent MRP initiative. PV holds a Methusalem Grant (BOF09/01M00709) from the Flemish Government. Research in the Fulda group is supported by the Deutsche Forschungsgemeinschaft, Federal Research Program IAP 6/18, European Research Program FP6 ApopTrain (MRTN-CT-035624) and FP7 Apo-Sys 200767.
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Vanlangenakker, N., Vanden Berghe, T., Bogaert, P. et al. cIAP1 and TAK1 protect cells from TNF-induced necrosis by preventing RIP1/RIP3-dependent reactive oxygen species production. Cell Death Differ 18, 656–665 (2011). https://doi.org/10.1038/cdd.2010.138
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DOI: https://doi.org/10.1038/cdd.2010.138
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