Abstract
Tight transcriptional regulation, alternative splicing and/or post-translational modifications of BH3-only proteins fine-tune their proapoptotic function. In this study, we characterize the gene locus of the BH3-only protein Bmf (Bcl-2-modifying factor) and describe the generation of two major isoforms from a common transcript in which initiation of protein synthesis involves leucine-coding CUG. BmfCUG and the originally described isoform, Bmf-short, display comparable binding affinities to prosurvival Bcl-2 family members, localize preferentially to the outer mitochondrial membrane and induce rapid Bcl-2-blockable apoptosis. Notably, endogenous Bmf expression is induced on forms of cell stress known to cause repression of the CAP-dependent translation machinery such as serum deprivation, hypoxia, inhibition of the PI3K/AKT pathway or mTOR, as well as direct pharmacological inhibition of the eukaryotic translation initiation factor eIF-4E. Knock down or deletion of Bmf reduces apoptosis under some of these conditions, demonstrating that Bmf can act as a sentinel for stress-impaired CAP-dependent protein translation machinery.
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Abbreviations
- Bmf:
-
Bcl-2-modifying factor
- BH:
-
Bcl-2 homology domain
- IVT:
-
in vitro transcription
- IVTT:
-
in vitro transcription and translation
- TSS:
-
transcription start site
- IRES:
-
intra-ribosomal entry site
- FF:
-
Firefly
- RN:
-
Renilla
- ECFP:
-
enhanced cyan fluorescent protein
- SV:
-
Simian virus
- MEF:
-
mouse embryonic fibroblast
- sh:
-
short hairpin
- ORF:
-
open reading frame
- eIF:
-
eukaryotic initiation factor
- DLC:
-
dynein light chain
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Acknowledgements
We thank Professors A Strasser, C Borner, M Schuler and W Doppler for cells and reagents; S Kiessling and F Müllauer for first experiments initiating this work; W Chmelewskij for help with MEF experiments; B Tomaselli for help with hypoxia experiments; Irene Gaggl for expert technical assistance, as well as all our colleagues in the lab for insightful discussions. This work was supported by grants from the AICR, St. Andrews, UK (no. 06-0440), the Graduate School for Molecular Biology and Oncology (MCBO) and the SFB021, both funded by the Austrian Science Fund (FWF).
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Grespi, F., Soratroi, C., Krumschnabel, G. et al. BH3-only protein Bmf mediates apoptosis upon inhibition of CAP-dependent protein synthesis. Cell Death Differ 17, 1672–1683 (2010). https://doi.org/10.1038/cdd.2010.97
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DOI: https://doi.org/10.1038/cdd.2010.97
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