title: Molecular basis for the sorting of the SNARE VAMP7 into endocytic clathrin-coated vesicles by the ArfGAP Hrb.
abstract: SNAREs provide the specificity and energy for the fusion of vesicles with their target membrane, but how they are sorted into the appropriate vesicles on post-Golgi trafficking pathways is largely unknown. We demonstrate that the clathrin-mediated endocytosis of the SNARE VAMP7 is directly mediated by Hrb, a clathrin adaptor and ArfGAP. Hrb wraps 20 residues of its unstructured C-terminal tail around the folded VAMP7 longin domain, demonstrating that unstructured regions of clathrin adaptors can select cargo. Disrupting this interaction by mutation of the VAMP7 longin domain or depletion of Hrb causes VAMP7 to accumulate on the cell's surface. However, the SNARE helix of VAMP7 binds back onto its longin domain, outcompeting Hrb for binding to the same groove and suggesting that Hrb-mediated endocytosis of VAMP7 occurs only when VAMP7 is incorporated into a cis-SNARE complex. These results elucidate the mechanism of retrieval of a postfusion SNARE complex in clathrin-coated vesicles.
authors.affiliation: Cambridge Institute for Medical Research and Department of Clinical Biochemistry, University of Cambridge, Addenbrooke's Hospital, Hills Road, Cambridge CB2 0XY, UK.
authors.forename: Paul R
authors.initials: PR
authors.lastname: Pryor
authors.forename: Lauren
authors.initials: L
authors.lastname: Jackson
authors.forename: Sally R
authors.initials: SR
authors.lastname: Gray
authors.forename: Melissa A
authors.initials: MA
authors.lastname: Edeling
authors.forename: Amanda
authors.initials: A
authors.lastname: Thompson
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Sanderson
authors.forename: Philip R
authors.initials: PR
authors.lastname: Evans
authors.forename: David J
authors.initials: DJ
authors.lastname: Owen
authors.forename: J Paul
authors.initials: JP
authors.lastname: Luzio
mesh_terms: D033942:Adaptor Proteins, Vesicular Transport / Q000737:chemistry / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D022163:Clathrin-Coated Vesicles / Q000378:metabolism*; D004705:Endocytosis; D006801:Humans; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D050683:R-SNARE Proteins / Q000737:chemistry* / Q000378:metabolism*; D020798:Two-Hybrid System Techniques
chemical_list: D033942:Adaptor Proteins, Vesicular Transport; D050683:R-SNARE Proteins; C432415:Sec22b protein, mouse; C496927:Sybl1 protein, mouse; C121241:VAMP7 protein, human
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