Abstract
In pathological conditions, the amount of DJ-1 determines whether a cell can survive or engage a cell death program. This is exemplified in epithelial cancers, in which DJ-1 expression is increased, while autosomal recessive early onset Parkinson’s disease mutations of DJ-1 generally lead to decreased stability and expression of the protein. We have shown previously that DJ-1 is cleaved by caspase-6 during induction of apoptosis. We demonstrate here that the N-terminal cleaved fragment of DJ-1 (DJ-1 Nt) is specifically expressed in the nucleus and promotes apoptosis in SH-SY5Y neuroblastoma cell lines. In addition, overexpression of DJ-1 Nt in different cell lines leads to a loss of clonogenic potential and sensitizes to staurosporin and 1-methyl-4-phenylpyridinium (MPP+)-mediated caspase activation and apoptosis. Importantly, inhibition of endogenous DJ-1 expression with sh-RNA or DJ-1 deficiency mimics the effect of DJ-1 Nt on cell growth and apoptosis. Moreover, overexpression of DJ-1 Nt increases reactive oxygen species (ROS) production, and sensitizes to MPP+-mediated apoptosis and DJ-1 oxidation. Finally, specific exclusion of DJ-1 Nt from the nucleus abrogates its pro-apoptotic effect. Taken together, our findings identify an original pathway by which generation of a nuclear fragment of DJ-1 through caspase 6-mediated cleavage induces ROS-dependent amplification of apoptosis.
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Abbreviations
- STS:
-
staurosporine
- MPP+:
-
1-methyl-4-phenylpyridinium
- EOPD:
-
early onset Parkinson’s disease
- PARP:
-
poly-ADP-ribose polymerase
- ROS:
-
reactive oxygen species
- DCFDA:
-
dichloro-dihydrofluorescein diacetate
- IP:
-
isoelectric point
- FCS:
-
fetal calf serum
- PVDF:
-
polyvinylidene difluoride
- CHAPS:
-
3-[(3–cholamidopropyl)dimethylammonio]-1-propanesulfonate
- PAGE:
-
polyacrylamide gel electrophoresis
- Ac-DEVD-AMC:
-
Ac-Asp-Glu-Val-Asp-AMC
- AMC:
-
7-amino-4-methylcoumarin
- S.D.:
-
standard deviation
- Z-VAD-fmk:
-
benzyloxycarbonyl-Val-Ala-Asp(OMe)-fluoromethylketone
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Acknowledgements
We are indebted to Professor Bill Earnshaw and Dr. Hiromi Ogawa for the kind gift of DT40 caspase 6+/+ and caspase 6−/− cells. This work was supported by grants from the Institut National des Cancéropoles (INCa PL2006-026 and PL2010-249) and by the Ligue Nationale contre le Cancer. GR is the recipient of fellowships from INCa and the Fondation de France. FC is supported by a grant from the Fondation pour la Recherche Médicale and by the Conseil General des Alpes-Maritimes. We acknowledge the C3M Imaging Core Facility (MICA) and genomic facility. We thank the Conseil Regional PACA and the Conseil Général des Alpes-Maritimes for their financial support to C3M.
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Robert, G., Puissant, A., Dufies, M. et al. The caspase 6 derived N-terminal fragment of DJ-1 promotes apoptosis via increased ROS production. Cell Death Differ 19, 1769–1778 (2012). https://doi.org/10.1038/cdd.2012.55
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DOI: https://doi.org/10.1038/cdd.2012.55
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