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Distinct endocytic pathways regulate TGF-β receptor signalling and turnover

Nature Cell Biology volume 5, pages 410–421 (2003)Cite this article

An Erratum to this article was published on 01 July 2003

Abstract

Endocytosis of cell surface receptors is an important regulatory event in signal transduction. The transforming growth factor β (TGF-β) superfamily signals to the Smad pathway through heteromeric Ser-Thr kinase receptors that are rapidly internalized and then downregulated in a ubiquitin-dependent manner. Here we demonstrate that TGF-β receptors internalize into both caveolin- and EEA1-positive vesicles and reside in both lipid raft and non-raft membrane domains. Clathrin-dependent internalization into the EEA1-positive endosome, where the Smad2 anchor SARA is enriched, promotes TGF-β signalling. In contrast, the lipid raft-caveolar internalization pathway contains the Smad7-Smurf2 bound receptor and is required for rapid receptor turnover. Thus, segregation of TGF-β receptors into distinct endocytic compartments regulates Smad activation and receptor turnover.

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Figure 1: TGF-β receptors localize to both Cav-1 and EEA1 positive compartments.
Figure 2: Dominant negative mutants of Dynamin or Eps15 modulate TGF-β receptor colocalization in subcellular compartments.
Figure 3: TGF-β receptor signal transduction and turnover is dependent on distinct subcellular compartments.
Figure 4: SARA and Smad7 (Smurf2) localize to distinct subcellular 0compartments.
Figure 5: Receptor binding proteins that associate with raft fractions stimulate receptor complex turnover.
Figure 6: SARA interferes with receptor complex degradation.
Figure 7: Sequestering of TGF-β receptors from lipid rafts permits signal transduction.

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Acknowledgements

The authors wish to thank S. Hearn (Mount Sinai Hospital, Toronto, EM facilities), J. Mui and J. Bergeron and A. Vali (Anatomy Cell Biology, McGill Montreal, EM Centre) and S. Doyle (Microscopy Imaging Laboratory, University of Toronto, EM facilities) for their assistance with the electron microscopy studies. The authors would also like to thank J. Bergeron, H. Benchabane, P. Cameron, R. Rasmussen and P. Kavsak for helpful advice, S. Kulkarni for help with deconvolution microscopy and L. Attisano and P. Cameron for critical review of the manuscript as well as L. Black, M. Pye and the members of the Wrana lab for assistance. This work was supported by grants to J. L. W. from the CIHR and the National Cancer Institute with funds from the Terry Fox run. C. L. R. is a Postdoctoral Fellow of the CIHR and J.L.W. is an International Investigator of the HHMI and an Investigator of the CIHR.

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  1. Gianni M. Di Guglielmo and Christine Le Roy: These authors contributed equally to this work.

Authors and Affiliations

  1. Programme in Molecular Biology and Cancer, Samuel Lunenfeld Research Institute, Mount Sinai Hospital, Toronto, Canada

    Gianni M. Di Guglielmo, Christine Le Roy, Anne F. Goodfellow & Jeffrey L. Wrana

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Correspondence to Jeffrey L. Wrana.

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Supplementary information

Supplementary Figures

Fig. S1. TGFβ receptors and biotinylated TGFβ co-localize to the EEA1 or Caveolin- 1 positive compartment. (PDF 12603 kb)

Fig. S2. TGFβ receptors do not co-localize with the late endosomal compartment, but partially co-localize with the recycling compartment.

Fig. S3. Ultrastructural and immunoelectron microscopy of TβRII.

Fig. S4. Comparison of transferrin and TGFβ ligand internalization.

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Di Guglielmo, G., Le Roy, C., Goodfellow, A. et al. Distinct endocytic pathways regulate TGF-β receptor signalling and turnover. Nat Cell Biol 5, 410–421 (2003). https://doi.org/10.1038/ncb975

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