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Hrs sorts ubiquitinated proteins into clathrin-coated microdomains of early endosomes

Nature Cell Biology volume 4, pages 394398 (2002) | Download Citation

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Abstract

After endocytosis, some membrane proteins recycle from early endosomes to the plasma membrane whereas others are transported to late endosomes and lysosomes for degradation1. Conjugation with the small polypeptide ubiquitin is a signal for lysosomal sorting2,3. Here we show that the hepatocyte growth factor-regulated tyrosine kinase substrate, Hrs4, is involved in the endosomal sorting of ubiquitinated membrane proteins. Hrs contains a clathrin-binding domain5, and by electron microscopy we show that Hrs localizes to flat clathrin lattices on early endosomes. We demonstrate that Hrs binds directly to ubiquitin by way of a ubiquitin-interacting motif (UIM), and that ubiquitinated proteins localize specifically to Hrs- and clathrin-containing microdomains. Whereas endocytosed transferrin receptors fail to colocalize with Hrs and rapidly recycle to the cell surface, transferrin receptors that are fused to ubiquitin interact with Hrs, localize to Hrs- and clathrin-containing microdomains and are sorted to the degradative pathway. Overexpression of Hrs strongly and specifically inhibits recycling of ubiquitinated transferrin receptors by a mechanism that requires a functional UIM. We conclude that Hrs sorts ubiquitinated membrane proteins into clathrin-coated microdomains of early endosomes, thereby preventing their recycling to the cell surface.

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Acknowledgements

We thank Eva Rønning for technical assistance and Philip Woodman for helpful advice. This work was supported by the Top Research Programme, the Research Council of Norway, the Norwegian Cancer Society, the Novo-Nordisk Foundation and the Anders Jahre's Foundation for the Promotion of Science.

Author information

Affiliations

  1. Department of Biochemistry, Institute for Cancer Research, the Norwegian Radium Hospital, Montebello, N-0310 Oslo, Norway stenmark@ulrik.uio.no

    • Camilla Raiborg
    • , Kristi G. Bache
    • , David J. Gillooly
    •  & Harald Stenmark
  2. Institute of Pathology, the National Hospital, N-0027 Oslo, Norway

    • Inger Helene Madshus
    •  & Espen Stang

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Competing interests

The authors declare no competing financial interests.

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    Figure S1

    Ubiquitination of TfR causes its sorting into the degradative endocytic pathway.

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DOI

https://doi.org/10.1038/ncb791

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