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VARP is a Rab32/38 effector that also binds to the endosomal/lysosomal R-SNARE VAMP7. VARP binding regulates VAMP7 participation in SNARE complex formation and can therefore influence VAMP7-mediated membrane fusion events. Mutant versions of VARP that cannot bind Rab32:GTP, designed on the basis of the VARP ankyrin repeat/Rab32:GTP complex structure described here, unexpectedly retain endosomal localization, showing that VARP recruitment is not dependent on Rab32 binding. We show that recruitment of VARP to the endosomal membrane is mediated by its direct interaction with VPS29, a subunit of the retromer complex, which is involved in trafficking from endosomes to the TGN and the cell surface. Transport of GLUT1 from endosomes to the cell surface requires VARP, VPS29, and VAMP7 and depends on the direct interaction between VPS29 and VARP. Finally, we propose that endocytic cycling of VAMP7 depends on its interaction with VARP and, consequently, also on retromer.

Original publication

DOI

10.1016/j.devcel.2014.04.010

Type

Journal article

Journal

Dev Cell

Publication Date

09/06/2014

Volume

29

Pages

591 - 606

Keywords

Amino Acid Sequence, Blotting, Western, Cell Membrane, Crystallography, X-Ray, Endosomes, Glucose Transporter Type 1, Guanine Nucleotide Exchange Factors, Guanosine Triphosphate, HeLa Cells, Humans, Immunoprecipitation, Molecular Sequence Data, Muscle Proteins, Mutagenesis, Site-Directed, Nuclear Proteins, Protein Binding, Protein Conformation, Protein Multimerization, Protein Transport, R-SNARE Proteins, Repressor Proteins, Sequence Homology, Amino Acid, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Vesicular Transport Proteins, rab GTP-Binding Proteins