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Apoptosis overrides survival signals through a caspase-mediated dominant-negative NF-κB loop

Nature Cell Biology volume 1, pages 227–233 (1999)Cite this article

Abstract

The transcription factor NF-κB is an important regulator of gene expression during immune and inflammatory responses, and can also protect against apoptosis. Here we show that endothelial cells undergo apoptosis when deprived of growth factors. Surviving viable cells exhibit increased activity of NF-κB, whereas apoptotic cells show caspase-mediated cleavage of the NF-κB p65/RelA subunit. This cleavage leads to loss of carboxy-terminal transactivation domains and a transcriptionally inactive p65 molecule. The truncated p65 acts as a dominant-negative inhibitor of NF-κB, promoting apoptosis, whereas an uncleavable, caspase-resistant p65 protects the cells from apoptosis. The generation of a dominant-negative fragment of p65 during apoptosis may be an efficient pro-apoptotic feedback mechanism between caspase activation and NF-κB inactivation.

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Figure 1: After deprivation of growth factors, NF-κB’s DNA-binding activity is induced in viable cells and is molecularly altered in apoptotic cells.
Figure 2: NF-κB transcriptional activity is induced in cells surviving growth-factor deprivation.
Figure 3: A proteolytic fragment of p65 appears during apoptosis induced by growth-factor deprivation.
Figure 4: p65 is a substrate for caspases in vitro and in vivo.
Figure 5: Identification of the caspase-cleavage sites in p65 and construction of uncleavable and truncation mutants.
Figure 6: Uncleavable p65 maintains κB-dependent transcription and rescues HUVECs from apoptosis.
Figure 7: The p65 truncation mutant inhibits the increase in κB-dependent transcription and survival induced by overexpression of wild-type p65.

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Acknowledgements

We thank U. Malyankar for helpful discussions and M. Almeida for technical assistance. This study was supported by NIH grant HL18645 (to R.R., E.W.R. and C.M.G.), National Science Foundation grant EEC9529161 (to C.M.G) and Innovative Medizinische Forschung grant LE 129835 (to B.L.).

Correspondence and requests for materials should be addressed to E.W.R.

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  1. Bodo Levkau and Marta Scatena: These authors contributed equally to this work

Authors and Affiliations

  1. Department of Pathology, University of Washington, Seattle, 98195-7470, Washington, USA

    Bodo Levkau, Marta Scatena, Cecilia M. Giachelli, Russell Ross & Elaine W. Raines

  2. Institute of Arteriosclerosis Research, University of Münster, Münster, 0-48149, Germany

    Bodo Levkau

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Correspondence to Elaine W. Raines.

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Levkau, B., Scatena, M., Giachelli, C. et al. Apoptosis overrides survival signals through a caspase-mediated dominant-negative NF-κB loop. Nat Cell Biol 1, 227–233 (1999). https://doi.org/10.1038/12050

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