Abstract
Death-associated protein kinase 1 (DAPK1) has been shown to have important roles in neuronal cell death in several model systems and has been implicated in multiple diseases, including Alzheimer’s disease (AD). However, little is known about the molecular mechanisms by which DAPK1 signals neuronal cell death. In this study, N-myc downstream-regulated gene 2 (NDRG2) was identified as a novel substrate of DAPK1 using phospho-peptide library screening. DAPK1 interacted with NDRG2 and directly phosphorylated the Ser350 residue in vitro and in vivo. Moreover, DAPK1 overexpression increased neuronal cell death through NDRG2 phosphorylation after ceramide treatment. In contrast, inhibition of DAPK1 by overexpression of a DAPK1 kinase-deficient mutant and small hairpin RNA, or by treatment with a DAPK1 inhibitor significantly decreased neuronal cell death, and abolished NDRG2 phosphorylation in cell culture and in primary neurons. Furthermore, NDRG2-mediated cell death by DAPK1 was required for a caspase-dependent poly-ADP-ribose polymerase cleavage. In addition, DAPK1 ablation suppressed ceramide-induced cell death in mouse brain and neuronal cell death in Tg2576 APPswe-overexpressing mice. Finally, levels of phosphorylated NDRG2 Ser350 and DAPK1 were significantly increased in human AD brain samples. Thus, phosphorylation of NDRG2 on Ser350 by DAPK1 is a novel mechanism activating NDRG2 function and involved in neuronal cell death regulation in vivo.
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Abbreviations
- AD:
-
Alzheimer’s disease
- DAPK1:
-
death-associated protein kinase 1
- NDRG2:
-
N-myc downstream-regulated gene 2
- Aβ:
-
amyloid-β oligomer
- WT:
-
wild-type
- KO:
-
knockout
- MEF:
-
mouse embryonic fibroblast
- CIP:
-
calf intestinal phosphatase
- PARP:
-
poly-ADP-ribose polymerase
- S.E.:
-
standard error
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Acknowledgements
We thank S-H Min for providing a NDRG2 construct and A Kimchi for providing DAPK1 KO mice. Human brain tissues were provided by the Neuropathology Core of the Massachusetts Alzheimer Disease Research Center (P50AG05134) and the Harvard Brain Tissue Resources Center (R24MH068855). The work was supported by grants from the Korea Healthcare Technology R&D Project, Ministry for Health & Welfare Affairs, Republic of Korea (HI08C2149) to BM Kim and NIH grant (R00AG033104), the Alzheimer’s Association (NIRG-12-258863), the American Federation for Aging Research, and the Massachusetts Alzheimer’s Disease Research Center (P50AG005134) to TH Lee.
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You, MH., Kim, B., Chen, CH. et al. Death-associated protein kinase 1 phosphorylates NDRG2 and induces neuronal cell death. Cell Death Differ 24, 238–250 (2017). https://doi.org/10.1038/cdd.2016.114
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DOI: https://doi.org/10.1038/cdd.2016.114
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