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p230 is associated with vesicles budding from the trans-Golgi network
P A Gleeson1, T J Anderson, J L Stow
1Department of Pathology and Immunology, Monash University Medical School, Melbourne, Victoria, Australia. pag@cobra.path.monash.edu.au
Journal of Cell Science
|December 1, 1996
Summary
Cytosolic protein p230 binds to Golgi membranes, regulated by G proteins. This protein is involved in forming specific non-clathrin-coated vesicles at the trans-Golgi network.
Area of Science:
- Cell Biology
- Molecular Biology
- Membrane Trafficking
Background:
- Transport vesicle formation necessitates cytosolic protein association with membranes.
- A brefeldin-A sensitive protein, p230, is found in the cytosol and associated with Golgi membranes.
Purpose of the Study:
- To investigate the localization of p230 on the trans-Golgi network (TGN).
- To determine the role of G protein activators in p230 binding to Golgi membranes and vesicle biogenesis.
Main Methods:
- Immunogold labeling of HeLa cell cryosections using compartment-specific markers.
- Treatment of permeabilized HeLa cells with GTPγS or AlF4-.
- Immunolabeling of isolated Golgi membranes.
- Dual immunogold labeling to distinguish p230 and p200 localization.
Main Results:
- p230 localizes to the TGN, with its binding to Golgi membranes enhanced by G protein activators (GTPγS, AlF4-).
- p230 is found on the cytoplasmic surface of TGN-derived budding structures and small vesicles, distinct from clathrin-coated vesicles.
- p230 and p200 localize to separate populations of TGN-derived vesicles, suggesting distinct vesicle types.
Conclusions:
- p230 is involved in the biogenesis of a specific population of non-clathrin-coated vesicles budding from the TGN.
- The recycling of p230 between the cytosol and TGN membranes is regulated by G proteins.
- Distinct populations of non-clathrin-coated vesicles are derived from the TGN.