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Acetylation-dependent regulation of endothelial Notch signalling by the SIRT1 deacetylase

Nature volume 473, pages 234–238 (2011)Cite this article

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Abstract

Notch signalling is a key intercellular communication mechanism that is essential for cell specification and tissue patterning, and which coordinates critical steps of blood vessel growth1,2,3 . Although subtle alterations in Notch activity suffice to elicit profound differences in endothelial behaviour and blood vessel formation2,3 , little is known about the regulation and adaptation of endothelial Notch responses. Here we report that the NAD+-dependent deacetylase SIRT1 acts as an intrinsic negative modulator of Notch signalling in endothelial cells. We show that acetylation of the Notch1 intracellular domain (NICD) on conserved lysines controls the amplitude and duration of Notch responses by altering NICD protein turnover. SIRT1 associates with NICD and functions as a NICD deacetylase, which opposes the acetylation-induced NICD stabilization. Consequently, endothelial cells lacking SIRT1 activity are sensitized to Notch signalling, resulting in impaired growth, sprout elongation and enhanced Notch target gene expression in response to DLL4 stimulation, thereby promoting a non-sprouting, stalk-cell-like phenotype. In vivo, inactivation of Sirt1 in zebrafish and mice causes reduced vascular branching and density as a consequence of enhanced Notch signalling. Our findings identify reversible acetylation of the NICD as a molecular mechanism to adapt the dynamics of Notch signalling, and indicate that SIRT1 acts as rheostat to fine-tune endothelial Notch responses.

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Figure 1: SIRT1 limits endothelial DLL4/Notch signalling and targets NICD for deacetylation.
Figure 2: Destabilization of NICD by SIRT1.
Figure 3: Inactivation of SIRT1 enhances endothelial Notch responses in mice.
Figure 4: Inactivation of SIRT1 leads to a cell-autonomous increase in Notch signalling and defective endothelial cell sprouting.

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Change history

  • 20 April 2011

    In Fig. 1, panel l was corrected; in Fig. 2, panels d, g, h and j were corrected.

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Acknowledgements

We are thankful to F. W. Alt, R. Kopan, Z. Lou, E. Seto, S. L. Berger, S. Diane Hayward, S. McMahon, G. Thurston and N. D. Lawson for reagents and to I. Dikic for comments. This work was supported by grants from the DFG (PO1306/1-1, SFB 834/A6 and Exc 147/1). F.D. was supported by the Interuniversity Attraction Poles Program–Belgian Science Policy (IUAP-BELSPO PVI/28). R.M. is supported by the Sidney Kimmel Cancer Research Foundation, a New Investigator Grant from the Massachusetts Life Sciences Center, an AFAR Research Grant and NIH grants (R01DK088190-01A1 and R01GM093072-01). H.G. is supported by Cancer Research UK, the European Molecular Biology Organisation Young Investigator Programme, and The Lister Institute of Preventive Medicine. H.G. and K.B. are supported by the Fondation Leducq Transatlantic Network of Excellence ARTEMIS. C.A.F. is supported by the Marie Curie FP7 People initiative. G.D. and M.M. thank F. Pezzimenti for fish care and technical help, and AIRC (Associazione Italiana per la Ricerca sul Cancro) for financial support.

Author information

Author notes

  1. Li-Kun Phng

    Present address: Present address: Cell Biology and Biophysics, European Molecular Biology Laboratory, D-69117 Heidelberg, Germany.,

  2. Gianluca Deflorian and Claudio A. Franco: These authors contributed equally to this work.

Authors and Affiliations

  1. Institute for Cardiovascular Regeneration, Centre of Molecular Medicine, Goethe University, D-60590 Frankfurt, Germany

    Virginia Guarani, Barbara Zimmermann, Stefanie Dimmeler & Michael Potente

  2. IFOM, the FIRC Institute of Molecular Oncology, IFOM-IEO Campus, 20139 Milan, Italy

    Gianluca Deflorian & Marina Mione

  3. Vascular Biology Laboratory, London Research Institute – Cancer Research UK, WC2A 3LY London, UK

    Claudio A. Franco, Li-Kun Phng, Katie Bentley & Holger Gerhardt

  4. Department of Cardiac Development and Remodeling, Max Planck Institute for Heart and Lung Research, D-61231 Bad Nauheim, Germany

    Marcus Krüger & Thomas Braun

  5. Laboratory of Protein Signaling and Interactions, GxABT, B-5030 Gembloux and Interdisciplinary Cluster for Applied Genoproteomics (GIGA-R), University of Liege, B-4000 Sart-Tilman, Belgium

    Louise Toussaint & Franck Dequiedt

  6. Massachusetts General Hospital Cancer Center, Harvard Medical School, Boston, 02114, Massachusetts, USA

    Raul Mostoslavsky

  7. Molecular Signal Transduction, Institute of Neurology (Edinger Institute), Goethe University, D-60590 Frankfurt, Germany

    Mirko H. H. Schmidt

  8. Vascular Research Centre, Institute for Cardiovascular Physiology, Goethe University, D-60590 Frankfurt, Germany

    Ralf P. Brandes

  9. Sirtris, a GSK Company, Cambridge, 02139, Massachusetts, USA

    Christoph H. Westphal

  10. Department of Cardiology, Internal Medicine III, Goethe University, D-60590 Frankfurt, Germany

    Andreas M. Zeiher & Michael Potente

  11. Consultant Group Leader, Vascular Patterning Laboratory, Vesalius Research Center, VIB, Campus Gasthuisberg, B-3000 Leuven, Belgium

    Holger Gerhardt

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Contributions

V.G. and M.P. designed and guided research. V.G., G.D., C.A.F., M.K., L.-K.P., K.B., L.T., F.D., M.H.H.S., B.Z., R.P.B., M.M., H.G. and M.P. performed research. V.G., G.D., C.A.F., M.K., L.-K.P., K.B., L.T., F.D., M.M., H.G. and M.P. analysed data. R.M., C.H.W. and T.B. provided reagents and/or technical support. T.B., A.M.Z. and S.D. gave conceptual advice. V.G., H.G. and M.P. wrote the paper. All authors commented on the manuscript.

Corresponding author

Correspondence to Michael Potente.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Figures

This file contains Supplementary Figures 1-16 with legends. (PDF 24705 kb)

Supplementary Movie 1

This movie shows simulation of a Sirt1 wild-type vessel. Tip cell selection occurs fast, movie represents 1 hour and 52 minutes (450 model time steps). Colour indicates NICD levels, purple - low, green – high (MOV 1098 kb)

Supplementary Movie 2

This movie shows simulation of a Sirt1-deficient vessel. Tip cell selection occurs slowly after initial oscillations in NICD levels. Colour indicates NICD levels, purple - low, green - high. (MOV 2410 kb)

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Guarani, V., Deflorian, G., Franco, C. et al. Acetylation-dependent regulation of endothelial Notch signalling by the SIRT1 deacetylase. Nature 473, 234–238 (2011). https://doi.org/10.1038/nature09917

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