Biochemical Requirements for the Targeting and Fusion of ER-Derived Transport Vesicles with Purified Yeast Golgi Membranes

In order for secretion to progress, ER-derived transport vesicles must target to, and fuse with the cis-Golgi compartment. These processes have been reconstituted using highly enriched membrane fractions and partially purified soluble components. The functionally active yeast Golgi membranes that ha...

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Bibliographic Details
Published in:The Journal of cell biology 1996-02, Vol.132 (3), p.277-289
Main Authors: Lupashin, Vladimir V., Hamamoto, Susan, Schekman, Randy W.
Format: Article
Language:English
Subjects:
Antibodies
Biochemistry
Biological Transport
Biomarkers
Cell Fractionation - methods
Cell membranes
Cells
Cellular biology
Cytosol
Endoplasmic Reticulum - physiology
Endoplasmic Reticulum - ultrastructure
Enzymes
Fungal Proteins - analysis
Golgi apparatus
Golgi Apparatus - physiology
Golgi Apparatus - ultrastructure
Intracellular Membranes - physiology
Intracellular Membranes - ultrastructure
Mannosyltransferases - analysis
Membrane Fusion
Membranes
Microscopy, Electron
Organelles - physiology
Organelles - ultrastructure
P branes
Proteins
Pyrophosphatases - analysis
Saccharomyces cerevisiae
Saccharomyces cerevisiae - genetics
Saccharomyces cerevisiae - physiology
Saccharomyces cerevisiae - ultrastructure
Transport vesicles
Yeast
Yeasts
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Description
Summary:In order for secretion to progress, ER-derived transport vesicles must target to, and fuse with the cis-Golgi compartment. These processes have been reconstituted using highly enriched membrane fractions and partially purified soluble components. The functionally active yeast Golgi membranes that have been purified are highly enriched in the cis-Golgi marker enzymes α1,6 mannosyltransferase and GDP-ase. Fusion of transport vesicles with these membranes requires both GTP and ATP hydrolysis, and depends on cytosolic and peripheral membrane proteins. At least two protein fractions from yeast cytosol are required for the reconstitution of ER-derived vesicle fusion. Soluble fractions prepared from temperature-sensitive mutants revealed requirements for the Ypt1p, Sec19p, Sly1p, Sec7p, and Uso1 proteins. A model for the sequential involvement of these components in the targeting and fusion reaction is proposed.
ISSN:0021-9525
1540-8140
DOI:10.1083/jcb.132.3.277