Wnt proteins are lipid-modified glycoproteins that play a central role in development, adult tissue homeostasis and disease. Secretion of Wnt proteins is mediated by the Wnt-binding protein Wntless (Wls), which transports Wnt from the Golgi network to the cell surface for release. It has recently been shown that recycling of Wls through a retromer-dependent endosome-to-Golgi trafficking pathway is required for efficient Wnt secretion, but the mechanism of this retrograde transport pathway is poorly understood. Here, we report that Wls recycling is mediated through a retromer pathway that is independent of the retromer sorting nexins SNX1–SNX2 and SNX5–SNX6. We have found that the unrelated sorting nexin, SNX3, has an evolutionarily conserved function in Wls recycling and Wnt secretion and show that SNX3 interacts directly with the cargo-selective subcomplex of the retromer to sort Wls into a morphologically distinct retrieval pathway. These results demonstrate that SNX3 is part of an alternative retromer pathway that functionally separates the retrograde transport of Wls from other retromer cargo.
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We thank C. Rabouille for critically reading the manuscript, M. Seaman for advice, D. Xue (University of Colorado, Boulder) for smIs34, M. Tabuchi (Kawasaki Medical School, Okayama, Japan) for the bacterial construct expressing 3×Flag-tagged retromer complex, S. Mitani (National Bioresource Project for the Nematode, Tokyo, Japan) for deletion mutants, A. Fire for expression vectors and the Caenorhabditis Genetic Center (University of Minnesota, Minneapolis) for strains. This work was funded by the Dutch Cancer Society (HUBR 2008-4114), the EU FP6 Programme Cells into Organs and NWO VIDI (016.076.317) (H.C.K.), an NWO VENI fellowship (M.J.L.), a Boehringer Ingelheim Foundation fellowship (M.H.), the Swiss National Science Foundation and the Forschungskredit of the University of Zürich (F.P. and K.B.) and the Wellcome Trust (089928/Z/09/Z and 085743) (P.J.C.). I.J.M. is a Wellcome Trust-funded PhD student (086777/Z/08/Z).
The authors declare no competing financial interests.
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Harterink, M., Port, F., Lorenowicz, M. et al. A SNX3-dependent retromer pathway mediates retrograde transport of the Wnt sorting receptor Wntless and is required for Wnt secretion. Nat Cell Biol 13, 914–923 (2011). https://doi.org/10.1038/ncb2281
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