Abstract
Cellular FLICE-inhibitory protein (c-FLIP) proteins are known as potent inhibitors of death receptor-mediated apoptosis by interfering with caspase-8 activation at the death-inducing signaling complex (DISC). Among the three human isoforms, c-FLIPlong, c-FLIPshort and c-FLIPR, the latter isoform is poorly characterized. We report here the characterization of murine c-FLIPR and show that it is the only short c-FLIP isoform expressed in mice. By generating several mutants, we demonstrate that both death effector domains (DEDs) are required for DISC binding and the antiapoptotic function of c-FLIPR. Surprisingly, the C-terminal tail is important for both protein stability and DISC recruitment. Three-dimensional modeling of c-FLIPR revealed a substantial similarity of the overall structures and potential interaction motifs with the viral FLIP MC159. We found, however, that c-FLIPR uses different structural motifs for its DISC recruitment. Whereas MC159 interferes with interaction and self-oligomerization of the DISC component FADD by its extensive hydrophilic surface, a narrow hydrophobic patch of c-FLIPR on the surface of DED2 is crucial for DISC association. Thus, despite the presence of similar tandem DEDs, viral and cellular FLIPs inhibit apoptosis by remarkably divergent mechanisms.
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Abbreviations
- CD95L:
-
CD95 ligand
- c-FLIP:
-
cellular FLICE-inhibitory protein
- CHX:
-
cycloheximide
- DED:
-
death effector domain
- DISC:
-
death-inducing signaling complex
- FADD:
-
Fas-associated death domain
- v-FLIP:
-
viral FLICE-inhibitory protein
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Acknowledgements
We thank Carina Meyer and Daniel Scholtyssik for expert technical assistance, and Dr Christian Schwerk for helpful discussions. We are also grateful to Drs Vishva Dixit, Thomas Hofmann, Michael Lenardo, Margot Thome and Harald Wajant for various reagents. This work was supported by grants from the Forschungskommission, Faculty of Medicine Düsseldorf, by the Deutsche Krebshilfe and the Deutsche Forschungsgemeinschaft (GK1033).
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Ueffing, N., Keil, E., Freund, C. et al. Mutational analyses of c-FLIPR, the only murine short FLIP isoform, reveal requirements for DISC recruitment. Cell Death Differ 15, 773–782 (2008). https://doi.org/10.1038/sj.cdd.4402314
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DOI: https://doi.org/10.1038/sj.cdd.4402314
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