Abstract
Budding of COP-coated vesicles (the likely carriers of newly synthesized proteins from the endoplasmic reticulum through the Golgi stack) from Golgi cisternae requires ADP-ribosylation factor (ARF), coatomer proteins from the cytosol, GTP, and fatty acyl-coenzyme A (CoA). The assembly of coated buds on the membranes requires coatomer, ARF, and GTP. When palmitoyl-CoA is added, membrane fission occurs at the coated bud, releasing coated vesicles. We show that COP-coated vesicles can be generated stepwise in vitro and isolated in a functionally active form, demonstrating that the minimal set of cytosolic components required for their formation as well as principal steps in their assembly have been identified.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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ADP-Ribosylation Factors
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Acyl Coenzyme A / metabolism
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Animals
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Biological Transport, Active
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CHO Cells
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Cell Compartmentation
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Coatomer Protein
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Cricetinae
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GTP-Binding Proteins / metabolism*
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Golgi Apparatus / metabolism*
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Guanosine Triphosphate / metabolism
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In Vitro Techniques
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Intracellular Membranes / metabolism*
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Membrane Glycoproteins / metabolism
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Membrane Proteins / metabolism*
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Microtubule-Associated Proteins / metabolism*
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Morphogenesis
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Organelles / metabolism
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Organelles / ultrastructure*
Substances
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Acyl Coenzyme A
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Coatomer Protein
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Membrane Glycoproteins
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Membrane Proteins
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Microtubule-Associated Proteins
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Guanosine Triphosphate
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GTP-Binding Proteins
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ADP-Ribosylation Factors