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d-Asb11 is an essential mediator of canonical Delta–Notch signalling

Abstract

In canonical Delta–Notch signalling, expression of Delta activates Notch in neighbouring cells, leading to downregulation of Delta in these cells1. This process of lateral inhibition results in selection of either Delta-signalling cells or Notch-signalling cells. Here we show that d-Asb11 is an important mediator of this lateral inhibition. In zebrafish embryos, morpholino oligonucleotide (MO)-mediated knockdown of d-Asb11 caused repression of specific Delta–Notch elements and their transcriptional targets, whereas these were induced when d-Asb11 was misexpressed. d-Asb11 also activated legitimate Notch reporters cell-non-autonomously in vitro and in vivo when co-expressed with a Notch reporter. However, it repressed Notch reporters when expressed in Delta-expressing cells. Consistent with these results, d-Asb11 was able to specifically ubiquitylate and degrade DeltaA both in vitro and in vivo. We conclude that d-Asb11 is a component in the regulation of Delta–Notch signalling, important in fine-tuning the lateral inhibition gradients between DeltaA and Notch through a cell non-autonomous mechanism.

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Figure 1: Delta-Notch and d-Asb11 function in a positive-feedback loop.
Figure 2: d-Asb11 transactivates specific Notch dependent reporters.
Figure 3: d-Asb11 binds to the Notch regulator Nrarp and the Notch ligand DeltaA.
Figure 4: d-Asb11 acts upstream of DeltaA.
Figure 5: Activation of Notch reporters by d-Asb11 requires other cells (a) When d-Asb11 was overexpressed in the same cell as the Notch reporter, Hes1 activity was elevated (left panel).

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Acknowledgements

We wish to thank J. A. Campos-Ortega for providing the MT–notch1a ICD construct, B. Appel for the MT–deltaA construct and A. Israel for the different Hes1–luciferase reporter constructs. We thank R. Dorsky, C. Houart, P. Ingham, J.P. Concordet for the riboprobes. We thank J. Lewis for DeltaD antibody, G. Strous for HA–Ubiquitin, U. Strähle for XDeltastu. We are indebted to R. Dorsky and R. Moon for providing TOPdGFP fish. We thank Y.J. Jiang for advice. We also wish to thank K. Österreicher and D. Kolmer (TissueGnostics, GmbH) for their help in the quantification of the co-localization studies described in Supplementary Information, Fig. S2. NWO Casimir, NWO Genomics, NWO ALW, TIPharma and the IAG program of the province of Groningen and the European Commission are thanked for financial support. We are especially grateful to S. van de Water, S. van den Brink, A. Visser and L. Glazenburg for technical assistance and to the animal facility of the Hubrecht Institute for care of zebrafish.

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S.H.D. contributed substantially to the conception and design of the study, acquisition, analysis and interpretation of the data, and drafting the manuscript; M.A.S.d.S. acquired, analysed and interpreted data; J.L.H., R.J.B., H.H.V., C.v.R. and A.B. acquired, analysed and interpreted data, and provided critical comments; M.P.P. analysed and interpreted the data; D.Z. acquired, analysed and interpreted data; M.P.P. and D.Z conceived and designed the study, and wrote and revised the manuscript.

Corresponding author

Correspondence to Maikel P. Peppelenbosch.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Information

Supplementary Figures S1, S2, S3, S4, S5, Supplementary Data and Supplementary Table 1 (PDF 805 kb)

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Diks, S., Sartori da Silva, M., Hillebrands, JL. et al. d-Asb11 is an essential mediator of canonical Delta–Notch signalling. Nat Cell Biol 10, 1190–1198 (2008). https://doi.org/10.1038/ncb1779

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