Formation of AP-3 transport intermediates requires Vps41 function

Nat Cell Biol. 1999 Oct;1(6):346-53. doi: 10.1038/14037.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adaptor Protein Complex alpha Subunits
  • Adaptor Proteins, Vesicular Transport
  • Alkaline Phosphatase / genetics
  • Alkaline Phosphatase / metabolism
  • Binding Sites
  • Biological Transport
  • Carrier Proteins / genetics
  • Carrier Proteins / metabolism*
  • Carrier Proteins / physiology
  • Cell Compartmentation
  • Cell Membrane / metabolism
  • Fungal Proteins / genetics
  • Fungal Proteins / metabolism*
  • Fungal Proteins / physiology
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism*
  • Monomeric Clathrin Assembly Proteins*
  • Mutagenesis
  • Nuclear Proteins*
  • Qa-SNARE Proteins
  • RNA-Binding Proteins / genetics
  • RNA-Binding Proteins / metabolism*
  • RNA-Binding Proteins / physiology
  • Saccharomyces cerevisiae Proteins*
  • Vacuoles / metabolism
  • Vesicular Transport Proteins*

Substances

  • Adaptor Protein Complex alpha Subunits
  • Adaptor Proteins, Vesicular Transport
  • Carrier Proteins
  • Fungal Proteins
  • Membrane Proteins
  • Monomeric Clathrin Assembly Proteins
  • NPL3 protein, S cerevisiae
  • Nuclear Proteins
  • Qa-SNARE Proteins
  • RNA-Binding Proteins
  • Saccharomyces cerevisiae Proteins
  • VAM3 protein, S cerevisiae
  • VPS41 protein, S cerevisiae
  • Vesicular Transport Proteins
  • clathrin assembly protein AP180
  • Alkaline Phosphatase