Nemo-like kinase suppresses notch signalling by interfering with formation of the notch active transcriptional complex

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. Access through your institution Buy or

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INHIBITION OF WNT SIGNALLING BY NOTCH VIA TWO DISTINCT MECHANISMS Article Open access 27 April 2021 NOTCH INTRACELLULAR DOMAINS FORM TRANSCRIPTIONALLY ACTIVE HETERODIMERIC COMPLEXES ON

SEQUENCE-PAIRED SITES Article Open access 02 January 2024 FUNCTIONAL ROLE OF THE FRIZZLED LINKER DOMAIN IN THE WNT SIGNALING PATHWAY Article Open access 05 May 2022 REFERENCES *

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neurogenesis. _Development_ 124, 693–702 (1997). CAS  PubMed  Google Scholar  Download references ACKNOWLEDGEMENTS We thank T. Honjo, T. C. Südhof, 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.). AUTHOR INFORMATION AUTHORS AND AFFILIATIONS * Unit on Nervous Development

Systems, 464-8602, Nagoya, Japan Tohru Ishitani, Tomoko Hirao, Miho Isoda & Motoyuki Itoh * Division of Biological Science, Group of Signal Transduction, Laboratory of Cell Regulation,

Graduate School of Science, 464-8602, Nagoya, Japan Tohru Ishitani, Maho Suzuki & Kunihiro Matsumoto * Institute for Advanced Research, Nagoya University, 464-8602, Nagoya, Japan

Motoyuki Itoh * Division of Cell Regulation Systems, Department of Post-Genome Science Center, Medical Institute of Bioregulation, Kyushu University, 812-8582, Fukuoka, Japan Tohru Ishitani 

& Shizuka Ishitani * Department of Molecular and Tumor Pathology, Chiba University Graduate School of Medicine, 260-8670, Chiba, Japan Kenichi Harigaya & Motoo Kitagawa Authors *

Tohru Ishitani View author publications You can also search for this author inPubMed Google Scholar * Tomoko Hirao View author publications You can also search for this author inPubMed 

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inPubMed Google Scholar * Shizuka Ishitani View author publications You can also search for this author inPubMed Google Scholar * Kenichi Harigaya View author publications You can also

search for this author inPubMed Google Scholar * Motoo Kitagawa View author publications You can also search for this author inPubMed Google Scholar * Kunihiro Matsumoto View author

publications You can also search for this author inPubMed Google Scholar * Motoyuki Itoh View author publications You can also search for this author inPubMed Google Scholar CONTRIBUTIONS

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.

CORRESPONDING AUTHORS Correspondence to Tohru Ishitani or Motoyuki Itoh. ETHICS DECLARATIONS COMPETING INTERESTS The authors declare no competing financial interests. SUPPLEMENTARY

INFORMATION SUPPLEMENTARY INFORMATION Supplementary Information (PDF 1585 kb) RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE 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 Download citation * Received: 13 October 2009 * Accepted: 14 December 2009 * Published: 31 January 2010 * Issue Date: March 2010 * DOI:

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