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Catalytic activity of the caspase-8–FLIPL complex inhibits RIPK3-dependent necrosis

Nature volume 471, pages 363–367 (2011)Cite this article

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Abstract

Caspase-8 has two opposing biological functions—it promotes cell death by triggering the extrinsic pathway of apoptosis, but also has a survival activity, as it is required for embryonic development1, T-lymphocyte activation2, and resistance to necrosis induced by tumour necrosis factor-α (TNF-α) and related family ligands3,4. Here we show that development of caspase-8-deficient mice is completely rescued by ablation of receptor interacting protein kinase-3 (RIPK3). Adult animals lacking both caspase-8 and RIPK3 display a progressive lymphoaccumulative disease resembling that seen with defects in CD95 or CD95-ligand (also known as FAS and FASLG, respectively), and resist the lethal effects of CD95 ligation in vivo. We have found that caspase-8 prevents RIPK3-dependent necrosis without inducing apoptosis by functioning in a proteolytically active complex with FLICE-like inhibitory protein long (FLIPL, also known as CFLAR), and this complex is required for the protective function.

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Figure 1: Casp8 −/− Ripk3 −/− mice are viable and functionally deficient for caspase-8.
Figure 2: Casp8 −/− Ripk3 −/− mice display progressive severe lymphoaccumulation.
Figure 3: Both catalytically active caspase-8 and FLIP are required for suppression of TNF-induced RIPK3-dependent death.
Figure 4: The caspase-8–FLIP heterodimeric complex suppresses RIPK3-dependent cell death.

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Acknowledgements

The authors thank W. J. Kaiser and E. S. Mocarski for their discussions and material support. We thank Ariad Pharmaceutical for providing the homo- and heterodimerization reagents. We thank the members of the St. Jude Immunology FACS core facility, as well as M. Johnson, L. Janke and the St. Jude Veterinary Pathology Core. We also thank the St. Jude Hartwell Center. This work was supported by NIH grant AI44828 to D.R.G. and CA69381 to G.S.S., as well as CIHR grant MOP 36537 to R.H. C.P.D. was supported by a fellowship grant from the Sass Foundation for Medical Research. This work was also supported by ALSAC.

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Authors and Affiliations

  1. Dept. of Immunology, St. Jude Children’s Research Hospital, Memphis, 38105, Tennessee, USA

    Andrew Oberst, Christopher P. Dillon, Ricardo Weinlich, Laura L. McCormick, Patrick Fitzgerald & Douglas R. Green

  2. Program in Apoptosis and Cell Death Research, Sanford-Burnham Medical Research Institute, La Jolla, 92037, California, USA

    Cristina Pop & Guy S. Salvesen

  3. Ontario Cancer Institute, University of Toronto, Toronto, Ontario M5G 2M9, Canada

    Razq Hakem

Authors
  1. Andrew Oberst
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  6. Cristina Pop
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Contributions

A.O. and D.R.G conceived the study and designed the experiments. C.P.D. and L.L.M. designed and conducted mouse breedings. C.P.D., R.W. and A.O. carried out all experiments involving mice and tissues from mice. A.O. and R.W. carried out experiments involving cell lines and produced western blots. P.F. provided essential logistical and administrative support. C.P. and G.S.S. conceived, designed and carried out in vitro dimerization assays and CrmA inhibition studies. R.H. produced the caspase-8flox animals that made the study possible.

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Correspondence to Douglas R. Green.

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Oberst, A., Dillon, C., Weinlich, R. et al. Catalytic activity of the caspase-8–FLIPL complex inhibits RIPK3-dependent necrosis. Nature 471, 363–367 (2011). https://doi.org/10.1038/nature09852

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