Abstract
Apoptosis represents an important cellular defence mechanism against viral pathogens by virtue of its ability to remove infected cells. Consequently, many viruses have developed numerous strategies to prevent or delay host cell apoptosis in order to achieve productive replication. Here we report that deletion of the F1L gene from the vaccinia genome results in increased apoptosis during infection. We demonstrate that F1L, which has no sequence homology to Bcl-2 family members, inhibits apoptosis at the level of mitochondria by binding to Bak. As a consequence, F1L prevents Bak activation, oligomerization and interaction with active Bax, all critical steps in the induction of apoptosis. We demonstrate that residues 64–84 of F1L interact directly with the Bcl-2 homology domain 3 (BH3) domain of Bak. This region of F1L has limited sequence similarity to known Bak-interacting BH3 domains. We also find that such additional BH3-like domains exist in the vaccinia genome. We conclude that F1L uses this specific, BH3-like domain to bind and inhibit Bak at the mitochondria.
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Abbreviations
- BH3 domain:
-
Bcl-2 homology domain 3
- STS:
-
staurosporine
- TM:
-
transmembrane
- WR:
-
vaccinia virus Western Reserve
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Acknowledgements
We thank D Hancock (Cancer Research UK, London, UK) for Bcl-2, Bcl-XL, Bax and Bak cDNAs, and Eyad Eddaoudi, Gary Warnes and Sabrina Leverrier for help and advice with the FACS analyses. We also thank Mike Mitchell for help with the FuzzPro analysis, Jacomine Krijnse-Locker for the anti-F17R antibody and Nicola O’Reilly for peptide synthesis and preparation of the peptide array. AP was supported by a Marie Curie postdoctoral fellowship (HPMF-CT-2002–01590). We have no conflicting financial interests.
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After submission of this paper, Wasilenko et al. reported that F1L binds Bak (J. Virol. 79, 14031, 2005).
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Postigo, A., Cross, J., Downward, J. et al. Interaction of F1L with the BH3 domain of Bak is responsible for inhibiting vaccinia-induced apoptosis. Cell Death Differ 13, 1651–1662 (2006). https://doi.org/10.1038/sj.cdd.4401853
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DOI: https://doi.org/10.1038/sj.cdd.4401853
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