Abstract
In the Drosophila wing, the Nedd4 ubiquitin ligases (E3s), dNedd4 and Su(dx), are important negative regulators of Notch signaling; they ubiquitinate Notch, promoting its endocytosis and turnover. Here, we show that Drosophila Nedd4 family interacting protein (dNdfip) interacts with the Drosophila Nedd4-like E3s. dNdfip expression dramatically enhances dNedd4 and Su(dx)-mediated wing phenotypes and further disrupts Notch signaling. dNdfip colocalizes with Notch in wing imaginal discs and with the late endosomal marker Rab7 in cultured cells. In addition, dNdfip expression in the wing leads to ectopic Notch signaling. Supporting this, expression of dNdfip suppressed Notch+/− wing phenotype and knockdown of dNdfip enhanced the Notch+/− wing phenotype. The increase in Notch activity by dNdfip is ligand independent as dNdfip expression also suppressed deltex RNAi and Serrate+/− wing phenotypes. The opposing effects of dNdfip expression on Notch signaling and its late endosomal localization support a model whereby dNdfip promotes localization of Notch to the limiting membrane of late endosomes allowing for activation, similar to the model previously shown with ectopic Deltex expression. When dNedd4 or Su(dx) are also present, dNdfip promotes their activity in Notch ubiquitination and internalization to the lysosomal lumen for degradation.
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Abbreviations
- Dx:
-
deltex
- Ser:
-
Serrate
- Su(dx):
-
Suppressor of deltex
- Wg:
-
wingless
- Ndfip:
-
Nedd4 family interacting protein
- dNdfip:
-
Drosophila Ndfip
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Acknowledgements
We thank S Hayashi, M Baron and T Shandala for the provision of fly stocks, C Wesley for the stably expressing Notch cell line S2N and A Paterson for performing some of the real-time PCR analysis. We also thank the Bloomington and VDRC Stock Centers, the Australian Drosophila Biomedical Research Support Facility (OzDros) for fly lines, and Developmental Studies Hybridoma Bank (University of Iowa) for antibodies. This work was funded by the National Health and Medical Research Council of Australia project grant (508085) and fellowship (399300) to SK.
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Dalton, H., Denton, D., Foot, N. et al. Drosophila Ndfip is a novel regulator of Notch signaling. Cell Death Differ 18, 1150–1160 (2011). https://doi.org/10.1038/cdd.2010.130
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DOI: https://doi.org/10.1038/cdd.2010.130
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