Abstract
HIAP2 is a multifunctional protein that is critically involved in the regulation of cell survival and apoptosis. Here, we show that HIAP2 5′ untranslated region functions as a strong inhibitor of translation. Sequence analysis of human, mouse and rat sequences revealed that there exists a short open reading frame (ORF) that is located just upstream of the HIAP2 coding sequence. The translation of this uORF severely inhibited translation of the downstream reporter gene in vivo but not in vitro. Point mutation that destroys the CUG initiating codon of uORF markedly enhanced translation of the reporter gene without affecting the mRNA levels. Our results identify a novel translational regulatory mechanism that controls the expression of HIAP2 and point to the importance of tight regulation of antiapoptotic gene expression.
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Abbreviations
- UTR:
-
untranslated region
- IAP:
-
inhibitor of apoptosis protein
- TNF:
-
tumour necrosis factor
- BIR:
-
baculoviral IAP repeat
- RING:
-
really interesting new gene
- IRES:
-
internal ribosome entry site
- CAT:
-
chloramphenicol acetyltransferase
- βgal:
-
β-galactosidase
- ELISA:
-
enzyme-linked immunosorbent assay
- RPA:
-
ribonuclease protection assay
- ONPG:
-
o-nitrophenyl-β-D-galactopyranoside
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Acknowledgements
We thank the members of our laboratory for stimulating discussions, Jaime Blais for the help with the MACS-VA500 microaerophilic workstation and the University of California San Francisco/Neurosurgery Tissue Bank for the SF-268 cell line. This work was supported by the operating grant from the Canadian Institutes of Health Research (# 43984). MH is a CIHR New Investigator.
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Warnakulasuriyarachchi, D., Ungureanu, N. & Holčík, M. The translation of an antiapoptotic protein HIAP2 is regulated by an upstream open reading frame. Cell Death Differ 10, 899–904 (2003). https://doi.org/10.1038/sj.cdd.4401256
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DOI: https://doi.org/10.1038/sj.cdd.4401256
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