Abstract
The SEC17 gene of Saccharomyces cerevisiae is required for vesicular transport between the endoplasmic reticulum and the Golgi apparatus. Here we report that the product of the SEC17 gene has the exact biochemical properties expected for a yeast homologue of the mammalian transport factor, alpha-SNAP. The DNA sequence of SEC17 codes for a protein of predicted molecular mass of 33 kDa. Immunoblotting indicates that Sec17p fractionates as a peripheral membrane protein and is mostly soluble when overexpressed, suggesting the presence of a saturable membrane receptor for Sec17p. Sec17p was purified from yeast cytosol using a SNAP-dependent in vitro mammalian Golgi transport assay. Kinetic analysis using this assay shows Sec17p acts temporally close to the fusion of transport vesicles with the medial Golgi compartment. In yeast extracts, Sec17p binds to Sec18p with a 1:1 stoichiometry. The interaction between Sec17p and Sec18p requires an activity provided by yeast membranes, and this putative membrane receptor activity is not extracted by high salt treatment of membranes.
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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Amino Acid Sequence
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Animals
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Base Sequence
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Blotting, Western
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Carrier Proteins / genetics*
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Carrier Proteins / physiology*
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Chromatography, Liquid
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Cloning, Molecular
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DNA, Fungal
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Electrophoresis, Polyacrylamide Gel
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Endoplasmic Reticulum / metabolism
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Fungal Proteins / genetics*
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Genes, Fungal
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Golgi Apparatus / metabolism
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Guanosine 5'-O-(3-Thiotriphosphate) / metabolism
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Kinetics
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Mammals
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Membrane Proteins / genetics*
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Membrane Proteins / physiology
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Molecular Sequence Data
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N-Ethylmaleimide-Sensitive Proteins
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Restriction Mapping
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Saccharomyces cerevisiae / genetics*
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Saccharomyces cerevisiae Proteins*
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Soluble N-Ethylmaleimide-Sensitive Factor Attachment Proteins
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Vesicular Transport Proteins*
Substances
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Carrier Proteins
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DNA, Fungal
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Fungal Proteins
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Membrane Proteins
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SEC17 protein, S cerevisiae
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Saccharomyces cerevisiae Proteins
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Soluble N-Ethylmaleimide-Sensitive Factor Attachment Proteins
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Vesicular Transport Proteins
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Guanosine 5'-O-(3-Thiotriphosphate)
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N-Ethylmaleimide-Sensitive Proteins
Associated data
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GENBANK/L01141
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GENBANK/M61455
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GENBANK/M80251
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GENBANK/M80491
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GENBANK/M80492
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GENBANK/M93104
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GENBANK/S96735
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GENBANK/S96741
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GENBANK/S96751
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GENBANK/S96754
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GENBANK/Z23261