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First published online 16 October 2007
doi: 10.1242/jcs.003343

Journal of Cell Science 120, 3850-3858 (2007)
Published by The Company of Biologists 2007
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Research Article

Clathrin-mediated endocytosis of a lipid-raft-associated protein is mediated through a dual tyrosine motif

Ruth Rollason1, Viktor Korolchuk1, Clare Hamilton1, Peter Schu2 and George Banting1,*

1 Department of Biochemistry, University of Bristol, Bristol, BS8 1T, UK
2 Zentrum für Biochemie und Molekulare Zellbiologie, Universität Goettingen, Germany

* Author for correspondence (e-mail: g.banting{at}bristol.ac.uk)

Accepted 17 August 2007

We have previously shown that the integral membrane protein CD317 has both a conventional transmembrane domain near its N-terminus and a C-terminal glycosyl-phosphatidylinositol (GPI) anchor. With the possible exception of a minor topological variant of the prion protein, there remain no other convincing examples of a mammalian protein with such a topology. CD317 is localised to cholesterol-rich lipid microdomains (`lipid rafts') in the plasma membrane and is internalised from the cell surface for delivery to a juxta-nuclear compartment (most probably the TGN). We have now investigated the mechanism by which CD317 is internalised and find that this raft-associated integral membrane protein is internalised through a clathrin-dependent pathway, internalisation is dependent upon a novel dual-tyrosine-based motif in the cytosolic domain of CD317, the cytosolic domain of CD317 can interact with the µ subunits of the AP2 and AP1 adaptor complexes, interaction with AP1 is required for delivery of CD317 back to the TGN, and removal of the GPI anchor from CD317 reduces the efficiency of CD317 internalisation. Collectively, these data indicate that CD317 is internalised and delivered back to the TGN by the sequential action of AP2 and AP1 adaptor complexes and that, surprisingly, the clathrin-mediated internalisation of CD317 occurs more efficiently if CD317 is localised to lipid rafts.

Key words: HM1.24, GPI, BST-2, Lipid raft, Endocytosis, B-cells


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