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A SNX3-dependent retromer pathway mediates retrograde transport of the Wnt sorting receptor Wntless and is required for Wnt secretion

Nature Cell Biology volume 13, pages 914–923 (2011)Cite this article

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Abstract

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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Figure 1: SNX3 is required for EGL-20 (Wnt) signalling and MIG-14 (Wls) recycling in C. elegans.
Figure 2: DSnx3 is required for Wg secretion and Wls recycling in the Drosophila wing imaginal disc.
Figure 3: Co-localization and physical interaction of SNX3 with the cargo-selective subcomplex of the retromer.
Figure 4: SNX3 co-localizes with Wls and facilitates membrane association of the cargo-selective subcomplex of the retromer.
Figure 5: Wls is contained within SNX3-positive vesicular carriers but is absent from SNX1 retromer-decorated tubular carriers.

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Acknowledgements

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).

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Author notes

  1. Fillip Port, Magdalena J. Lorenowicz and Ian J. McGough: These authors contributed equally to this work

  2. Marie Silhankova is a Present address: Department of Cell Biology, Faculty of Science, Charles University Prague, Vinicna 7, 128 00 Prague 2, Czech Republic

  3. Peter J. Cullen and Hendrik C. Korswagen: Joint senior authors

Authors and Affiliations

  1. Hubrecht Institute, Royal Netherlands Academy of Arts and Sciences and University Medical Center Utrecht, Uppsalalaan 8, 3584 CT, Utrecht, The Netherlands

    Martin Harterink, Magdalena J. Lorenowicz, Marie Silhankova, Marco C. Betist, Roy G. H. P. van Heesbeen, Teije C. Middelkoop & Hendrik C. Korswagen

  2. Institute of Molecular Life Sciences, University of Zurich, Winterthurerstrasse 190, CH-8057 Zurich, Switzerland

    Fillip Port & Konrad Basler

  3. Department of Biochemistry, Henry Wellcome Integrated Signaling Laboratories, School of Medical Sciences, University of Bristol, Bristol BS8 1TD, UK

    Ian J. McGough, Jan R. T. van Weering & Peter J. Cullen

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Contributions

M.H., M.S., T.C.M., M.C.B., R.G.H.P.H. and H.C.K. designed and carried out the C. elegans experiments, F.P. and K.B. designed and carried out the Drosophila experiments, M.J.L., I.J.M., J.R.T.W., H.C.K. and P.J.C. designed and carried out the cell biological analysis of SNX3 function in tissue culture cells and M.H., K.B., P.J.C. and H.C.K. wrote the paper.

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Correspondence to Peter J. Cullen or Hendrik C. Korswagen.

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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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