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Formation of AP-3 transport intermediates requires Vps41 function
P Rehling1, T Darsow, D J Katzmann
1Division of Cellular and Molecular Medicine and Howard Hughes Medical Institute, University of California, San Diego, La Jolla, California 92093-0668, USA.
Nature Cell Biology
|November 24, 1999
Summary
The AP-3 adaptor protein complex is crucial for transporting proteins to the yeast vacuole. Vps41 protein interacts with AP-3, indicating a joint role in forming transport intermediates at the late Golgi.
Area of Science:
- Cell Biology
- Molecular Biology
- Protein Transport
Background:
- Transport of specific membrane proteins to the yeast vacuole depends on the AP-3 adaptor protein complex.
- Understanding the molecular mechanisms of this vesicular transport pathway is essential.
Purpose of the Study:
- To biochemically analyze the formation and cargo content of AP-3 transport intermediates.
- To elucidate the molecular requirements for AP-3 mediated vesicular transport to the yeast vacuole.
Main Methods:
- Utilized a vam3tsf mutant yeast strain to accumulate AP-3 transport intermediates.
- Isolated and analyzed accumulated membrane vesicles using biochemical methods and immunocytochemistry.
- Investigated the role of AP-3 and Vps41 in intermediate formation.
Main Results:
- Accumulated vesicles in the vam3tsf mutant contained AP-3 adaptors and alkaline phosphatase (ALP) pathway cargoes.
- Inactivation of AP-3 or Vps41 inhibited the formation of these vesicular intermediates.
- Demonstrated that Vps41 binds to the AP-3 delta-adaptin subunit.
Conclusions:
- Vesicular transport to the yeast vacuole relies on the AP-3 adaptor protein complex.
- Vps41 and AP-3 function together in forming ALP pathway transport intermediates at the late Golgi.
- Vps41's interaction with AP-3 delta-adaptin is key to this process.