Abstract
The cellular level of the CDC25A phosphatase is tightly regulated during both the normal and genotoxic-perturbed cell cycle. Here, we describe a caspase-dependent cleavage of this protein at residue D223 in non-genotoxic apoptotic conditions. This specific proteolysis generates a catalytically active C-terminal fragment that localizes to the nuclear compartment. Accumulation of this active CDC25A fragment leads to reduced inhibitory phosphorylation of the CDC25A substrate cyclin-dependent kinase 2 (CDK2) on Tyr15. Moreover, CDK2 was found stably associated with this fragment, as well as with an ectopically expressed CDC25A224–525 truncation mutant that mimicks the cleavage product. Ectopic expression of this mutant induced CDK2 Tyr15 dephosphorylation, whereas its catalytically inactive version did not. Finally, this 224–525 mutant initiated apoptosis when transfected into HeLa cells, whereas its catalytic inactive form did not. Altogether, this study demonstrates for the first time that caspase-dependent cleavage of CDC25A is a central step linking CDK2 activation with non-genotoxic apoptotic induction.
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Abbreviations
- AML:
-
acute myeloid leukemia
- CDK2:
-
cyclin-dependent kinase 2
- FACS:
-
fluorescence-activated cell sorting
- FDP:
-
fluorescein diphosphate
- NES:
-
nuclear export sequence
- NLS:
-
nuclear localization sequence
- PARP:
-
poly (ADP-ribose) polymerase
- STS:
-
staurosporin
- TNF-α:
-
tumor necrosis factor alpha
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Acknowledgements
We are very grateful to Professor E Solary (Dijon, France) for providing us with the U937-p35 cell line. We also thank Fatima L'Faqihi-Olive for expert assistance with the FACS experiments. This study was supported by the Institut National de la Santé et de la Recherche Médicale, by the Association Laurette Fugain (Grant no. R05027BB), by the Association pour la Recherche sur le Cancer (ARC, Grant no. 3638) by the ‘Ligue Nationale contre le Cancer’ (Equipe labellisée 2005 to BD) and by the Institut National du Cancer (INCA, Grant no. PL103).
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Mazars, A., Fernandez-Vidal, A., Mondesert, O. et al. A caspase-dependent cleavage of CDC25A generates an active fragment activating cyclin-dependent kinase 2 during apoptosis. Cell Death Differ 16, 208–218 (2009). https://doi.org/10.1038/cdd.2008.142
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DOI: https://doi.org/10.1038/cdd.2008.142
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