Abstract
Apoptosis is mediated by cysteine-dependent, aspartate-directed proteases of the caspase family that proteolyse strategic intracellular substrates to induce cell suicide. We describe here that engagement of apoptotic processes by Fas triggering or by staurosporine stimulation leads to the caspase-dependent inactivation of the nuclear factor kappa B (NF-κB) pathway after cleavage of IKK1 (IκB kinase 1) and NEMO (NF-κB essential modulator), which are needed to transduce NF-κB activation signals. In this study, we have analyzed in more detail, the role of NEMO cleavage, as NEMO, but not IKK1, is important for the pro-survival actions of NF-κB. We demonstrate that NEMO is cleaved after Asp355 to remove the last 64 C-terminal amino acids. This short form was unable to rescue NF-κB activation by tumor necrosis factor-α (TNF-α) when transfected in NEMO-deficient cells. Consequently, inactivation of NEMO resulted in an inhibition of the expression of antiapoptotic NF-κB-target genes coding for caspase inhibitors (cIAP-1, cIAP-2) or adaptors of the TNF receptor family. NEMO-deficient Jurkat cells transiently expressing a non-cleavable mutant of NEMO were less sensitive to TNF-α-induced apoptosis. Therefore, downmodulation of NF-κB activation via the proteolytic cleavage of NEMO could represent an amplification loop for apoptosis.
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Abbreviations
- EMSA:
-
electrophoretic mobility shift assay
- IκB:
-
inhibitor of kappa B
- IAP:
-
inhibitor of apoptosis
- IKK:
-
I kappa B kinase
- NEMO:
-
NF-κB essential modulator
- NF-κB:
-
nuclear factor kappa B
- PARP:
-
poly(ADP ribose) polymerase
- PMA:
-
phorbol myristate acetate
- RPA:
-
ribonuclease protection assay
- STS:
-
staurosporine
- TNF-α:
-
tumor necrosis factor
- TRAF:
-
TNF receptor associated factor
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Acknowledgements
We appreciate the critical reading of the manuscript by Dr. Robert Herrington (Toronto, Canada). We thank Drs. M Dreano (Merck-Serono International, Geneva, Switzerland), J Blenis (Boston, USA) and JC Chambard (CNRS, Nice, France) for the kind gift of AS602868, caspase-8 deficient Jurkat variants, active caspase-8 cDNA respectively. This work is supported by INSERM and grants from ARC (3392) and la Fondation de France, comité Leucémies.
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Frelin, C., Imbert, V., Bottero, V. et al. Inhibition of the NF-κB survival pathway via caspase-dependent cleavage of the IKK complex scaffold protein and NF-κB essential modulator NEMO. Cell Death Differ 15, 152–160 (2008). https://doi.org/10.1038/sj.cdd.4402240
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DOI: https://doi.org/10.1038/sj.cdd.4402240
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