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Notch post-translationally regulates β-catenin protein in stem and progenitor cells

Nature Cell Biology volume 13, pages 1244–1251 (2011)Cite this article

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Abstract

Cellular decisions of self-renewal or differentiation arise from integration and reciprocal titration of numerous regulatory networks. Notch and Wnt/β-catenin signalling often intersect in stem and progenitor cells and regulate each other transcriptionally. The biological outcome of signalling through each pathway often depends on the context and timing as cells progress through stages of differentiation. Here, we show that membrane-bound Notch physically associates with unphosphorylated (active) β-catenin in stem and colon cancer cells and negatively regulates post-translational accumulation of active β-catenin protein. Notch-dependent regulation of β-catenin protein did not require ligand-dependent membrane cleavage of Notch or the glycogen synthase kinase- 3β-dependent activity of the β-catenin destruction complex. It did, however, require the endocytic adaptor protein Numb and lysosomal activity. This study reveals a previously unrecognized function of Notch in negatively titrating active β-catenin protein levels in stem and progenitor cells.

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Figure 1: Notch negatively regulates active β-catenin in stem cells independently of RBP-J.
Figure 2: Notch1 negatively regulates active β-catenin in ESCs and physically interacts with β-catenin.
Figure 3: Membrane-bound Notch1 negatively regulates active β-catenin levels through Numb and Numb-like in stem cells.
Figure 4: Notch-mediated degradation of β-catenin requires Numb and lysosomal activity.
Figure 5: GSIs negatively regulate Wnt signalling and cell expansion in colon cancer cells by blocking Notch cleavage.

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Acknowledgements

We thank R. Kopan (Washington University), M. Nakafuku (Cincinnati Children’s Hospital), T. Honjo (Kyoto University) and P. Stanley (Albert Einstein College of Medicine) for providing tethered Notch constructs, Notch deletion constructs, RBP-Jflox mice and Notch1lbd/lbd ES cells, respectively. The authors thank G. Howard and S. Ordway for editorial assistance, Srivastava and Kwon laboratory members for discussions, B. Taylor for assistance with manuscript and figure preparation and B. Bruneau for critical reading of the manuscript. We also thank J. Fish and C. Miller in the Gladstone Histology core. C.K. was supported by grants from the American Heart Association Beginning Grant-in-Aid and National Heart, Lung, and Blood Institute/National Institutes of Health (NHLBI/NIH; 1K99HL092234, 4R00HL09223); I.N.K. was supported by a March of Dimes Basil O’Connor Award; D.S. was supported by grants from NHLBI/NIH (P01 HL089707, U01 HL100406), the California Institute for Regenerative Medicine and the Younger Family Foundation. This work was supported by NIH/National Center for Research Resources grant C06 RR018928 to the Gladstone Institute.

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Author notes

  1. Vishal Nigam

    Present address: Present address: Division of Cardiology, Department of Pediatrics, University of California, San Diego 92903, USA,

  2. Chulan Kwon and Paul Cheng: These authors contributed equally to this work

Authors and Affiliations

  1. Gladstone Institute of Cardiovascular Disease and Departments of Pediatrics and Biochemistry & Biophysics, University of California, San Francisco, 1650 Owens Street, San Francisco, California 94158, USA

    Chulan Kwon, Paul Cheng, Isabelle N. King, Vishal Nigam & Deepak Srivastava

  2. Division of Cardiology, Department of Medicine, Johns Hopkins University, 720 Rutland Avenue, Ross 954B, Baltimore, Maryland 21205, USA

    Chulan Kwon, Peter Andersen & Lincoln Shenje

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Contributions

C.K. designed, carried out and supervised in vivo and in vitro work and wrote the manuscript. P.C. designed and carried out in vivo and in vitro work and wrote the manuscript. I.N.K. carried out Notch Co-IP and western analyses. P.A. cultured embryonic stem cells and carried out luciferase assays. L.S. carried out immunocytochemistry and confocal microscopy. V.N. isolated mesenchymal stem cells and carried out western analyses. D.S. designed and supervised this work and wrote the manuscript.

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Correspondence to Chulan Kwon or Deepak Srivastava.

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Competing interests

D.S. is a scientific co-founder of iPierian Inc. and is a member of the Scientific Advisory Board of iPierian Inc. and RegeneRx Pharmaceuticals.

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Kwon, C., Cheng, P., King, I. et al. Notch post-translationally regulates β-catenin protein in stem and progenitor cells. Nat Cell Biol 13, 1244–1251 (2011). https://doi.org/10.1038/ncb2313

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