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

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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.

Change history

  • 20 April 2011

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


  1. 1

    Kopan, R. & Ilagan, M. X. The canonical Notch signaling pathway: unfolding the activation mechanism. Cell 137, 216–233 (2009)

  2. 2

    Roca, C. & Adams, R. H. Regulation of vascular morphogenesis by Notch signaling. Genes Dev. 21, 2511–2524 (2007)

  3. 3

    Phng, L. K. & Gerhardt, H. Angiogenesis: a team effort coordinated by notch. Dev. Cell 16, 196–208 (2009)

  4. 4

    Milne, J. C. et al. Small molecule activators of SIRT1 as therapeutics for the treatment of type 2 diabetes. Nature 450, 712–716 (2007)

  5. 5

    Kurooka, H. & Honjo, T. Functional interaction between the mouse Notch1 intracellular region and histone acetyltransferases PCAF and GCN5. J. Biol. Chem. 275, 17211–17220 (2000)

  6. 6

    Oswald, F. et al. p300 acts as a transcriptional coactivator for mammalian Notch-1. Mol. Cell. Biol. 21, 7761–7774 (2001)

  7. 7

    Kim, M. Y. et al. Tip60 histone acetyltransferase acts as a negative regulator of Notch1 signaling by means of acetylation. Mol. Cell. Biol. 27, 6506–6519 (2007)

  8. 8

    Potente, M. et al. SIRT1 controls endothelial angiogenic functions during vascular growth. Genes Dev. 21, 2644–2658 (2007)

  9. 9

    Hellström, M. et al. Dll4 signalling through Notch1 regulates formation of tip cells during angiogenesis. Nature 445, 776–780 (2007)

  10. 10

    Ridgway, J. et al. Inhibition of Dll4 signalling inhibits tumour growth by deregulating angiogenesis. Nature 444, 1083–1087 (2006)

  11. 11

    Suchting, S. et al. The Notch ligand Delta-like 4 negatively regulates endothelial tip cell formation and vessel branching. Proc. Natl Acad. Sci. USA 104, 3225–3230 (2007)

  12. 12

    Lobov, I. B. et al. Delta-like ligand 4 (Dll4) is induced by VEGF as a negative regulator of angiogenic sprouting. Proc. Natl Acad. Sci. USA 104, 3219–3224 (2007)

  13. 13

    Phng, L. K. et al. Nrarp coordinates endothelial Notch and Wnt signaling to control vessel density in angiogenesis. Dev. Cell 16, 70–82 (2009)

  14. 14

    Benedito, R. et al. The notch ligands Dll4 and Jagged1 have opposing effects on angiogenesis. Cell 137, 1124–1135 (2009)

  15. 15

    Bentley, K., Mariggi, G., Gerhardt, H. & Bates, P. A. Tipping the balance: robustness of tip cell selection, migration and fusion in angiogenesis. PLoS Comput. Biol. 5, e1000549 (2009)

  16. 16

    Siekmann, A. F. & Lawson, N. D. Notch signalling limits angiogenic cell behaviour in developing zebrafish arteries. Nature 445, 781–784 (2007)

  17. 17

    Leslie, J. D. et al. Endothelial signalling by the Notch ligand Delta-like 4 restricts angiogenesis. Development 134, 839–844 (2007)

  18. 18

    Jakobsson, L. et al. Heparan sulfate in trans potentiates VEGFR-mediated angiogenesis. Dev. Cell 10, 625–634 (2006)

  19. 19

    Jakobsson, L. et al. Endothelial cells dynamically compete for the tip cell position during angiogenic sprouting. Nature Cell Biol. 12, 943–953 (2010)

  20. 20

    Picard, F. et al. Sirt1 promotes fat mobilization in white adipocytes by repressing PPAR-gamma. Nature 429, 771–776 (2004)

  21. 21

    Hisahara, S. et al. Histone deacetylase SIRT1 modulates neuronal differentiation by its nuclear translocation. Proc. Natl Acad. Sci. USA 105, 15599–15604 (2008)

  22. 22

    Takata, T. & Ishikawa, F. Human Sir2-related protein SIRT1 associates with the bHLH repressors HES1 and HEY2 and is involved in HES1- and HEY2-mediated transcriptional repression. Biochem. Biophys. Res. Commun. 301, 250–257 (2003)

  23. 23

    Prozorovski, T. et al. Sirt1 contributes critically to the redox-dependent fate of neural progenitors. Nature Cell Biol. 10, 385–394 (2008)

  24. 24

    Kitamura, T. et al. A Foxo/Notch pathway controls myogenic differentiation and fiber type specification. J. Clin. Invest. 117, 2477–2485 (2007)

  25. 25

    Donmez, G., Wang, D., Cohen, D. E. & Guarente, L. SIRT1 suppresses β-amyloid production by activating the alpha-secretase gene ADAM10. Cell 142, 320–332 (2010)

  26. 26

    Finkel, T., Deng, C. X. & Mostoslavsky, R. Recent progress in the biology and physiology of sirtuins. Nature 460, 587–591 (2009)

  27. 27

    Potente, M. et al. Involvement of Foxo transcription factors in angiogenesis and postnatal neovascularization. J. Clin. Invest. 115, 2382–2392 (2005)

  28. 28

    North, B. J. et al. The human Sir2 ortholog, SIRT2, is an NAD+-dependent tubulin deacetylase. Mol. Cell 11, 437–444 (2003)

  29. 29

    Kim, J. E., Chen, J. & Lou, Z. DBC1 is a negative regulator of SIRT1. Nature 451, 583–586 (2008)

  30. 30

    Shevchenko, A. et al. In-gel digestion for mass spectrometric characterization of proteins and proteomes. Nature Protocols 1, 2856–2860 (2006)

  31. 31

    Rappsilber, J., Mann, M. & Ishihama, Y. Protocol for micro-purification, enrichment, pre-fractionation and storage of peptides for proteomics using StageTips. Nature Protocols 2, 1896–1906 (2007)

  32. 32

    Olsen, J. V. et al. A dual pressure linear ion trap Orbitrap instrument with very high sequencing speed. Mol. Cell. Proteomics 8, 2759–2769 (2009)

  33. 33

    Cox, J. & Mann, M. MaxQuant enables high peptide identification rates, individualized p.p.b.-range mass accuracies and proteome-wide protein quantification. Nature Biotechnol. 26, 1367–1372 (2008)

  34. 34

    Vintersten, K. et al. Mouse in red: red fluorescent protein expression in mouse ES cells, embryos, and adult animals. Genesis 40, 241–246 (2004)

  35. 35

    Cheng, H. L. et al. Developmental defects and p53 hyperacetylation in Sir2 homolog (SIRT1)-deficient mice. Proc. Natl Acad. Sci. USA 100, 10794–10799 (2003)

  36. 36

    Lawson, N. D. & Weinstein, B. M. In vivo imaging of embryonic vascular development using transgenic zebrafish. Dev. Biol. 248, 307–318 (2002)

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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

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.

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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