期刊
CELLULAR SIGNALLING
卷 28, 期 4, 页码 246-254出版社
ELSEVIER SCIENCE INC
DOI: 10.1016/j.cellsig.2016.01.003
关键词
Non-canonical NOTCH signaling; Delta-like 1 homolog (Dlk1); Tissue development; Two mammalian hybrid assay
类别
资金
- Novo Nordisk Foundation
- Danish National Research Council [09-073648]
- Lundbeck Foundation [R48-A4785, R112-A9634]
- Laegeforeningen [2011-3271/480853-109]
- Tomrermester Alfred Andersen og Hustru's Fond
- Hertha Christensens Foundation
- Eva Fraenkel Foundation
- A.P. Moller Foundation
- SPAR Nord
- Dep. of Clinical Biochemistry and Pharmacology/Odense University Hospital
- Henry Fraenkel Foundation
Canonical NOTCH signaling, known to be essential for tissue development, requires the Delta-Serrate-LAG2 (DSL) domain for NOTCH to interact with its ligand. However, despite lacking DSL, Delta-like 1 homolog (DLK1), a protein that plays a significant role in mammalian development, has been suggested to interact with NOTCH1 and act as an antagonist. This non-canonical interaction is, however controversial, and evidence for a direct interaction, still lacking in mammals. In this study, we elucidated the putative DLK1-NOTCH1 interaction in a mammalian context. Taking a global approach and using Dlk1(+/+) and Dlkl(-/-) mouse tissues at E16.5, we demonstrated that several NOTCH signaling pathways indeed are affected by DLK1 during tissue development, and this was supported by a lower activation of NOTCH1 protein in Dlk1(+/+) embryos. Likewise, but using a distinct Dlk1-manipulated (siRNA) setup in a mammalian cell line, NOTCH signaling was substantially inhibited by DLK1. Using a mammalian two-hybrid system, we firmly established that the effect of DLK1 on NOTCH signaling was due to a direct interaction between DLK1 and NOTCH1. By careful dissection of this mechanism, we found this interaction to occur between EGF domains 5 and 6 of DLK1 and EGF domains 10-15 of NOTCH1. Thus, our data provide the first evidence for a direct interaction between DLK1 and NOTCH1 in mammals, and substantiate that non-canonical NOTCH ligands exist, adding to the complexity of NOTCH signaling. (C) 2016 Elsevier Inc All rights reserved.
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