Modular mechanism of Wnt signaling inhibition by Wnt inhibitory factor 1


Wnt morphogens control embryonic development and homeostasis in adult tissues. In vertebrates the N-terminal WIF domain (WIF-1WD) of Wnt inhibitory factor 1 (WIF-1) binds Wnt ligands. Our crystal structure of WIF-1WD reveals a previously unidentified binding site for phospholipid; two acyl chains extend deep into the domain, and the head group is exposed to the surface. Biophysical and cellular assays indicate that there is a WIF-1WD Wnt-binding surface proximal to the lipid head group but also implicate the five epidermal growth factor (EGF)-like domains (EGFs I–V) in Wnt binding. The six-domain WIF-1 crystal structure shows that EGFs I–V are wrapped back, interfacing with WIF-1WD at EGF III. EGFs II–V contain a heparan sulfate proteoglycan (HSPG)-binding site, consistent with conserved positively charged residues on EGF IV. This combination of HSPG- and Wnt-binding properties suggests a modular model for the localization of WIF-1 and for signal inhibition within morphogen gradients.

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Figure 1: The WIF-1WD-EGF-I structure and lipid-binding cavity.
Figure 2: The contributions of the WIF and EGF-like domains to Wnt inhibition.
Figure 3: A discontinuous Wnt-binding site on the WIF domain.
Figure 4: The structure of WIF-1ΔC.
Figure 5: HSPG binding contributes to a modular mechanism for WIF-1 function.

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We thank the staff of the European Synchrotron Radiation Facility and Diamond Light Source for assistance with data collection; T.S. Walter and K. Harlos for help with crystallization; B.M. Kessler and K. di Gleria for MS analysis; B.J.C. Janssen for help with solvent flattening; and E. Seiradake, Y. Zhao and M.A. Jones for help with tissue culture. T.M. and E.Y.J. are funded by Cancer Research UK, C.S. by the Wellcome Trust and A.R.A. by the UK Medical Research Council.

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T.M., A.R.A., C.S. and E.Y.J. designed the project. T.M. performed all the experiments. W.L. contributed to WIF-1 protein expression. A.R.A. and C.S. contributed to X-ray data collection and analysis. T.M., A.R.A., C.S. and E.Y.J. analyzed the data and wrote the manuscript.

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Correspondence to Christian Siebold or E Yvonne Jones.

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Supplementary Methods and Supplementary Figures 1–9 (PDF 1879 kb)

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Malinauskas, T., Aricescu, A., Lu, W. et al. Modular mechanism of Wnt signaling inhibition by Wnt inhibitory factor 1. Nat Struct Mol Biol 18, 886–893 (2011).

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