Abstract
A unified model for initiator caspase activation has previously been proposed based on the biochemical analysis of caspase-8 and -9. Caspase-2 is structurally related to caspase-9, but its mechanism of activation is not known. Using an uncleavable mutant of caspase-2, we show that dimerization (and not processing) is the key event that drives initial procaspase-2 activation. Following dimerization, caspase-2 undergoes autocatalytic cleavage that promotes its stable dimerization and further enhances the catalytic activity of caspase-2. Although the caspase-2 zymogen does not require cleavage for the initial acquisition of activity, intersubunit cleavage is required to generate levels of activity required to induce cell death by overexpression. We also provide evidence that the reported disulfide bond linkage between two caspase-2 monomers is dispensable for caspase-2 dimerization. As caspase-2 does not require cleavage for its initial activation, our findings confirm caspase-2 to be a bona fide initiator caspase.
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Abbreviations
- CARD:
-
caspase recruitment domain
- GST:
-
glutathione S-transferase
- PAGE:
-
polyacrylamide gel electrophoresis
- PCR:
-
polymerase chain reaction
- WT:
-
wild type
- IVT:
-
in vitro translated
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Acknowledgements
We thank members of our laboratory and Stuart Pitson for the supply of reagents and helpful discussions. This work was supported by the National Health and Medical Research Council of Australia, and the Cancer Council of South Australia.
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Baliga, B., Read, S. & Kumar, S. The biochemical mechanism of caspase-2 activation. Cell Death Differ 11, 1234–1241 (2004). https://doi.org/10.1038/sj.cdd.4401492
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DOI: https://doi.org/10.1038/sj.cdd.4401492
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