Regulation of endosomal membrane traffic by a Gadkin/AP-1/kinesin KIF5 complex.
Endosomes and endosomal vesicles (EVs) rapidly move along cytoskeletal filaments allowing them to exchange proteins and lipids between different endosomal compartments, lysosomes, the trans-Golgi network (TGN), and the plasma membrane. The precise mechanisms that connect membrane traffic between the TGN and perinuclear endosomal compartments with motor-protein driven transport have largely remained elusive. Here we show that Gadkin (also termed gamma-BAR), a peripheral membrane protein localized to the TGN and to TGN-derived EVs, directly associates with the clathrin adaptor AP-1 and with the motor protein kinesin KIF5, thereby potentially regulating EV dynamics. Gadkin overexpression induced the dispersion of transferrin (Tf)- and Rab4-positive EVs to the cell periphery, whereas KIF5B-depleted cells displayed a perinuclear concentration. Functional experiments suggest that the role of Gadkin as a regulator of endosomal membrane traffic critically depends on complex formation with both AP-1 and KIF5. Our data thus provide a direct molecular link between TGN-derived EVs and the microtubule-based cytoskeleton.
Schmidt
Michael R
MR
Institute of Chemistry and Biochemistry, Department of Membrane Biochemistry, Freie Universität and Charité Universitätsmedizin Berlin, Takustrasse 6, 14195 Berlin, Germany.
Maritzen
Tanja
T
Kukhtina
Viktoria
V
Higman
Victoria A
VA
Doglio
Laura
L
Barak
Naomi N
NN
Strauss
Holger
H
Oschkinat
Hartmut
H
Dotti
Carlos G
CG
Haucke
Volker
V
D000818:Animals; D001693:Biological Transport, Active / Q000502:physiology; D019556:COS Cells; D002522:Chlorocebus aethiops; D002846:Chromatography, Affinity; D011992:Endosomes / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D047468:Immunoprecipitation; D016547:Kinesins / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D046912:Multiprotein Complexes / Q000378:metabolism*; D018808:Transcription Factor AP-1 / Q000378:metabolism*
C500885:AP1AR protein, human; C500506:KIF5A protein, human; D008565:Membrane Proteins; D046912:Multiprotein Complexes; D018808:Transcription Factor AP-1; D016547:Kinesins
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