Abstract
The Notch signalling pathway has a crucial function in determining cell fates in multiple tissues within metazoan organisms1. On binding to ligands, the Notch receptor is cleaved proteolytically and releases its intracellular domain (NotchICD). The NotchICD enters the nucleus and acts cooperatively with other factors to stimulate the transcription of target genes. High levels of Notch-mediated transcriptional activation require the formation of a ternary complex consisting of NotchICD, CSL (CBF-1, suppressor of hairless, LAG-1) and a Mastermind family member2,3,4,5. However, it is still not clear how the formation of the ternary complex is regulated. Here we show that Nemo-like kinase (NLK) negatively regulates Notch-dependent transcriptional activation by decreasing the formation of this ternary complex. Using a biochemical screen, we identified Notch as a new substrate of NLK. NLK-phosphorylated Notch1ICD is impaired in its ability to form a transcriptionally active ternary complex. Furthermore, knockdown of NLK leads to hyperactivation of Notch signalling and consequently decreases neurogenesis in zebrafish. Our results both define a new function for NLK and reveal a previously unidentified mode of regulation in the Notch signalling pathway.
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Acknowledgements
We thank T. Honjo, T. C. SĂĽdhof, U. Lendahl, K. Yasutomo, C. Kintner, D. Hayward, J. D. Griffin, L. Wu, M. Takeichi, S. Chiba, A. Kikuchi, H. Fujisawa and K. Hozumi for providing plasmid vectors, antibody and cultured cells; H. Matsuo for technical assistance; members of the H. Aiba laboratory (especially T. Sunohara) for technical advice; and J. Ninomiya-Tsuji for helpful discussions. This research was supported by the Yamada Science Foundation (T.I.), the Astellas Foundation for Research on Metabolic Disorders (T.I. and M.I.), the Program for Improvement of Research Environment for Young Researchers from SCF commissioned by MEXT of Japan (T.I. and M.I.), and the Grants-in-Aid for Scientific Research programs in Japan (T.I., K.M. and M.I.).
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T.I. designed the research, performed most of the experiments, analysed data and wrote the paper. T.H., M.S., M. Isoda and S.I. performed experiments and analysed data. K.H. participated in discussions and helped write the paper. M.K. designed the research and performed the experiments, analysed data and wrote the paper. K.M. designed the initial experiments to identify NLK substrates, participated discussions, and helped write the paper. M. Itoh designed the project and experiments, supervised the research, coordinated experiments and wrote the paper.
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Ishitani, T., Hirao, T., Suzuki, M. et al. Nemo-like kinase suppresses Notch signalling by interfering with formation of the Notch active transcriptional complex. Nat Cell Biol 12, 278–285 (2010). https://doi.org/10.1038/ncb2028
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DOI: https://doi.org/10.1038/ncb2028
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