Abstract
The repulsive guidance molecule (RGM) is a membrane-bound protein that has diverse functions in the developing central nervous system. Identification of neogenin as a receptor for RGM provided evidence of its cell death-inducing activity in the absence of RGM. Here, we show that the serine/threonine kinase death-associated protein kinase (DAPK) is involved in the signal transduction of neogenin. Neogenin interacts with DAPK and reduces DAPK autophosphorylation on Ser308 in vitro. Neogenin-induced cell death is abolished in the presence of RGM or by blocking DAPK. Although neogenin overexpression or RGM downregulation in the chick neural tube in vivo induces apoptosis, coexpression of the dominant-negative mutant or small-interference RNA of DAPK attenuates this proapoptotic activity. Thus, RGM/neogenin regulates cell fate by controlling the DAPK activity.
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Abbreviations
- RGM:
-
repulsive guidance molecule
- CNS:
-
central nervous system
- DAPK:
-
death-associated protein kinase
- DCC:
-
deleted in colorectal cancer
- siRNA:
-
small-interference RNA
- TUNEL:
-
terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling
- CaM:
-
calmodulin
- ICD:
-
intracellular domain
- HEK:
-
human embryonic kidney
- CGN:
-
cerebellar granule neuron
- HH:
-
Hamburger Hamilton
- MLC:
-
myosin light chain
- GFP:
-
green fluorescent protein
- DMEM:
-
Dulbecco's modified Eagle's medium
- FBS:
-
fetal bovine serum
- PFA:
-
paraformaldehyde
- BSA:
-
bovine serum albumin
- DAPI:
-
4′,6′-diamidino-2-phenylindole
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Acknowledgements
This work was supported by a Research Grant from the National Institute of Biomedical Innovation (05-12) and Grant-in-Aid for Young Scientists (S) from JSPS. We thank Tetsuichiro Saito and Yuko Muroyama for technical guidance for in ovo electroporation.
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Fujita, Y., Taniguchi, J., Uchikawa, M. et al. Neogenin regulates neuronal survival through DAP kinase. Cell Death Differ 15, 1593–1608 (2008). https://doi.org/10.1038/cdd.2008.92
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DOI: https://doi.org/10.1038/cdd.2008.92
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