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USP4 is regulated by AKT phosphorylation and directly deubiquitylates TGF-β type I receptor

Abstract

The stability and membrane localization of the transforming growth factor-β (TGF-β) type I receptor (TβRI) determines the levels of TGF-β signalling. TβRI is targeted for ubiquitylation-mediated degradation by the SMAD7–SMURF2 complex. Here we performed a genome-wide gain-of-function screen and identified ubiquitin-specific protease (USP) 4 as a strong inducer of TGF-β signalling. USP4 was found to directly interact with TβRI and act as a deubiquitylating enzyme, thereby controlling TβRI levels at the plasma membrane. Depletion of USP4 mitigates TGF-β-induced epithelial to mesenchymal transition and metastasis. Importantly, AKT (also known as protein kinase B), which has been associated with poor prognosis in breast cancer, directly associates with and phosphorylates USP4. AKT-mediated phosphorylation relocates nuclear USP4 to the cytoplasm and membrane and is required for maintaining its protein stability. Moreover, AKT-induced breast cancer cell migration was inhibited by USP4 depletion and TβRI kinase inhibition. Our results uncover USP4 as an important determinant for crosstalk between TGF-β and AKT signalling pathways.

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Figure 1: USP4 is a DUB required for TGF-β signalling.
Figure 2: USP4 interacts with TβRI and deubiquitylates TβRI.
Figure 3: USP4 stabilizes membrane receptor levels of TβRI.
Figure 4: USP4 increases TGF-β-induced EMT, invasion and metastasis.
Figure 5: AKT phosphorylates USP4.
Figure 6: AKT activation promotes the membrane and cytoplasmic localization of USP4.
Figure 7: AKT-mediated phosphorylation affects USP4 stability and DUB activity towards TβRI.

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Acknowledgements

We are grateful to M. van Dinther for performing the 125I–TGF-β labelling experiment, A. Teunisse and A. G. Jochemsen (Leiden University Medical Center, Leiden, The Netherlands) for 14-3-3 expression plasmids and purification of GST 14-3-3 fusion proteins, and H. van Dam for critical reading of the manuscript. We thank K. Iwata, S. Lin and S. Piccolo for reagents. This work was supported by a Netherlands Organization of Scientific Research grant (MW-NWO 918.66.606) and the Centre for Biomedical Genetics.

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L.Z., F.Z. and P.t.D. conceived and designed the experiments; L.Z. and F.Z. carried out the biochemical and functional assays in cells. Y.D., with help from E.S.-J., performed the zebrafish assays. R.G. and H.H. conducted the zebrafish embryo assays, C.M., K-A.S. and C.X.L. performed the genome-wide gain-of-function screen and J.A.P. and C.X.L. analysed the results. L.Z., F.Z. and P.t.D. wrote the manuscript.

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Correspondence to FangFang Zhou or Peter ten Dijke.

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Zhang, L., Zhou, F., Drabsch, Y. et al. USP4 is regulated by AKT phosphorylation and directly deubiquitylates TGF-β type I receptor. Nat Cell Biol 14, 717–726 (2012). https://doi.org/10.1038/ncb2522

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