Formation of AP-3 transport intermediates requires Vps41 function.
Transport of a subset of membrane proteins to the yeast vacuole requires the function of the AP-3 adaptor protein complex. To define the molecular requirements of vesicular transport in this pathway, we used a biochemical approach to analyse the formation and content of the AP-3 transport intermediate. A vam3tsf (vacuolar t-SNARE) mutant blocks vesicle docking and fusion with the vacuole and causes the accumulation of 50-130-nanometre membrane vesicles, which we isolated and showed by biochemical analysis and immunocytochemistry to contain both AP-3 adaptors and alkaline phosphatase (ALP) pathway cargoes. Inactivation of AP-3 or the protein Vps41 blocks formation of this vesicular intermediate. Vps41 binds to the AP-3 delta-adaptin subunit, suggesting that they function together in the formation of ALP pathway transport intermediates at the late Golgi.
Rehling
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P
Division of Cellular and Molecular Medicine and Howard Hughes Medical Institute, University of California, San Diego, La Jolla, California 92093-0668, USA.
Darsow
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Katzmann
D J
DJ
Emr
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SD
D033965:Adaptor Protein Complex alpha Subunits; D033942:Adaptor Proteins, Vesicular Transport; D000469:Alkaline Phosphatase / Q000235:genetics / Q000378:metabolism; D001665:Binding Sites; D001692:Biological Transport; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D002451:Cell Compartmentation; D002462:Cell Membrane / Q000378:metabolism; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D034041:Monomeric Clathrin Assembly Proteins; D016296:Mutagenesis; D009687:Nuclear Proteins; D050765:Qa-SNARE Proteins; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D029701:Saccharomyces cerevisiae Proteins; D014617:Vacuoles / Q000378:metabolism; D033921:Vesicular Transport Proteins
D033965:Adaptor Protein Complex alpha Subunits; D033942:Adaptor Proteins, Vesicular Transport; D002352:Carrier Proteins; D005656:Fungal Proteins; D008565:Membrane Proteins; D034041:Monomeric Clathrin Assembly Proteins; C077734:NPL3 protein, S cerevisiae; D009687:Nuclear Proteins; D050765:Qa-SNARE Proteins; D016601:RNA-Binding Proteins; D029701:Saccharomyces cerevisiae Proteins; C107706:VAM3 protein, S cerevisiae; C083878:VPS41 protein, S cerevisiae; D033921:Vesicular Transport Proteins; C439023:clathrin assembly protein AP180; D000469:Alkaline Phosphatase