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SNX27 mediates retromer tubule entry and endosome-to-plasma membrane trafficking of signalling receptors

Nature Cell Biology volume 13, pages 715–721 (2011)Cite this article

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Abstract

Endocytic sorting of signalling receptors between recycling and degradative pathways is a key cellular process controlling the surface complement of receptors and, accordingly, the cell’s ability to respond to specific extracellular stimuli. The β2 adrenergic receptor (β2AR) is a prototypical seven-transmembrane signalling receptor that recycles rapidly and efficiently to the plasma membrane after ligand-induced endocytosis. β2AR recycling is dependent on the receptor’s carboxy-terminal PDZ ligand and Rab4 (refs 1, 2). This active sorting process is required for functional resensitization of β2AR-mediated signalling3,4. Here we show that sequence-directed sorting occurs at the level of entry into retromer tubules and that retromer tubules are associated with Rab4. Furthermore, we show that sorting nexin 27 (SNX27) serves as an essential adaptor protein linking β2ARs to the retromer tubule. SNX27 does not seem to directly interact with the retromer core complex, but does interact with the retromer-associated Wiskott–Aldrich syndrome protein and SCAR homologue (WASH) complex. The present results identify a role for retromer in endocytic trafficking of signalling receptors, in regulating a receptor-linked signalling pathway, and in mediating direct endosome-to-plasma membrane traffic.

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Figure 1: Rapid recycling β2ARs selectively enter retromer-associated endosomal tubules.
Figure 2: Knockdown of retromer by RNAi inhibits β2AR recycling and misroutes internalized β2ARs to lysosomes.
Figure 3: Retromer depletion preferentially affects β2ARs over TFRs traversing the same endosomes.
Figure 4: β2AR and CIMPR follow divergent trafficking paths on exiting from the same retromer-associated tubule.
Figure 5: SNX27 serves as an adaptor for β2AR, sorting it into the retromer tubule.

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Acknowledgements

We thank B. Padilla, J. Bonifacino, Y. Prabhu, N. Gulbahce, S. Duleh, M. Welch, R. Rojas, J. Hislop, M. Puthenveedu, K. Mostov, J. Weissman, K. Thorn, the Nikon Imaging Center, A. Burlingame and the UCSF Mass Spectrometry facility (supported by P41RR001614) for reagents, advice, technical training and support. Last, we thank H. Bourne and M. Ray for critical readings of the manuscript. This work was supported by research grants from the National Institutes of Health. P.T. was supported by the National Science Foundation. N.J.K. is a Searle and Keck Young Investigator Fellow.

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Authors and Affiliations

  1. Department of Psychiatry, University of California at San Francisco, San Francisco, California 94158, USA

    Paul Temkin, Ben Lauffer & Mark von Zastrow

  2. Department of Cellular and Molecular Pharmacology, University of California at San Francisco, San Francisco, California 94158, USA

    Stefanie Jäger & Nevan J. Krogan

  3. Department of Bioengineering and Therapeutic Sciences, University of California at San Francisco, San Francisco, California 94158, USA

    Peter Cimermancic

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Contributions

P.T. and M.v.Z. conceived the project and wrote the manuscript. P.T. carried out most of the experiments with contributions from B.L. Mass spectrometry was carried out by S.J., analysed by P.C., in the laboratory of N.J.K.

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Correspondence to Mark von Zastrow.

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Temkin, P., Lauffer, B., Jäger, S. et al. SNX27 mediates retromer tubule entry and endosome-to-plasma membrane trafficking of signalling receptors. Nat Cell Biol 13, 715–721 (2011). https://doi.org/10.1038/ncb2252

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