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Tankyrase inhibition stabilizes axin and antagonizes Wnt signalling

Abstract

The stability of the Wnt pathway transcription factor β-catenin is tightly regulated by the multi-subunit destruction complex. Deregulated Wnt pathway activity has been implicated in many cancers, making this pathway an attractive target for anticancer therapies. However, the development of targeted Wnt pathway inhibitors has been hampered by the limited number of pathway components that are amenable to small molecule inhibition. Here, we used a chemical genetic screen to identify a small molecule, XAV939, which selectively inhibits β-catenin-mediated transcription. XAV939 stimulates β-catenin degradation by stabilizing axin, the concentration-limiting component of the destruction complex. Using a quantitative chemical proteomic approach, we discovered that XAV939 stabilizes axin by inhibiting the poly-ADP-ribosylating enzymes tankyrase 1 and tankyrase 2. Both tankyrase isoforms interact with a highly conserved domain of axin and stimulate its degradation through the ubiquitin-proteasome pathway. Thus, our study provides new mechanistic insights into the regulation of axin protein homeostasis and presents new avenues for targeted Wnt pathway therapies.

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Figure 1: XAV939 inhibits Wnt/β-catenin signalling by increasing axin protein levels.
Figure 2: Identification of the cellular efficacy targets of XAV939.
Figure 3: Tankyrase modulates axin protein levels.
Figure 4: Tankyrase physically and functionally interacts with axin.
Figure 5: XAV939 stabilizes axin protein and inhibits ubiquitination of axin.
Figure 6: XAV939 inhibits DLD-1 colony formation in an axin-dependent manner.

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Acknowledgements

We thank D. Patel, F. Harbinski, J. Leighton-Davies, R. de Beaumont, X. Xiang, K. Bean, C. Xin, S. Zhao, B. Zhang and M. Xu for technical assistance, G. Wussler, H. Urquiza and W. Dai for zebrafish maintenance, and I. Cornella Taracido, S. Cleaver, A. Hernandez and Y. Ben-Neriah for comments and advice. In addition we are indebted to B. Kuster, J. Rick, M. Raida and A. Scholten for continued support and discussion.

Author Contributions S.-M.A.H., A.C., F.St., G.A.M., E.W., V.M., S.F., C.Lu, D.C., M.W.K., C.Le., P.M.F., J.A.T., T.B., J.A.P., A.B. and F.C. conceived and designed the study. S.-M.A.H., Y.M.M., S.L., A.C., F.St., G.A.M., O.C., E.W., Y.Z., S.W., M.H., X.S., C.W., C.M., A.F., R.T., F.Se., W.S., H.C., M.Sh., C.R., M.Sc., J.S., S.G., A.B. and F.C. designed and implemented experiments. S.-M.A.H., F.St., A.B., Y.M.M. and F.C. wrote the paper.

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Correspondence to Feng Cong.

Supplementary information

Supplementary Information

This file contains Supplementary Methods, Supplementary Figures S1-S9 with Legends, and a Legend for Table S1 (see separate file s2). (PDF 2084 kb)

Supplementary Table 1

This file contains the competition data for all 699 identified proteins (see file s1 for full Legend). (XLS 98 kb)

Supplementary Methods

This file contains the methods for the synthesis of XAV939. (PDF 88 kb)

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Huang, SM., Mishina, Y., Liu, S. et al. Tankyrase inhibition stabilizes axin and antagonizes Wnt signalling. Nature 461, 614–620 (2009). https://doi.org/10.1038/nature08356

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