A rat brain Sec1 homologue related to Rop and UNC18 interacts with syntaxin.
Sec1 is a hydrophilic protein that plays an essential role in exocytosis from the yeast Saccharomyces cerevisiae. Two high copy suppressors of mutations in the Sec1 gene, SSO1 and SSO2, were recently identified that encode proteins of the syntaxin family. Syntaxin (a T-SNARE), together with SNAP-25 and synaptobrevin/VAMP (a T- and a V-SNARE, respectively), is thought to form the core of the docking-fusion complex in synaptic vesicle exocytosis. Proteins that exhibit similarity to Sec1 were identified in the nervous system of Drosophila melanogaster (Rop) and Caenorhabditis elegans (UNC18). Based on the amino acid sequence alignment of Sec1, Rop, and UNC18, we have used a PCR-based approach to isolate a rat brain cDNA encoding a Sec1 homologue. The cDNA hybridizes to a 3.5-kb brain-specific mRNA by Northern blot analysis and encodes a protein of 593 amino acids (rbSec1). Antibodies raised against a central portion of rbSec1 recognize a 67.5-kDa protein in total homogenates of rat brain but not of nonneuronal tissues. When incubated with a Triton X-100 brain extract, rbSec1-glutathione S-transferase (GST) fusion protein, but not GST protein alone, specifically interacts with syntaxin but not with SNAP-25 or synaptobrevin/VAMP. We conclude that the function of proteins of the Sec1 family in membrane fusion involves an interaction with a T-SNARE.
Garcia
E P
EP
Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06510.
Gatti
E
E
Butler
M
M
Burton
J
J
De Camilli
P
P
D000595:Amino Acid Sequence; D000818:Animals; D000954:Antigens, Surface / Q000235:genetics / Q000378:metabolism*; D015152:Blotting, Northern; D001921:Brain / Q000378:metabolism*; D029742:Caenorhabditis elegans Proteins; D002352:Carrier Proteins; D029721:Drosophila Proteins; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D015801:Helminth Proteins / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D051938:Munc18 Proteins; D009154:Mutation; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism*; D010750:Phosphoproteins; D016133:Polymerase Chain Reaction; D051381:Rats; D017386:Sequence Homology, Amino Acid; D050827:Syntaxin 1; D033921:Vesicular Transport Proteins
D000954:Antigens, Surface; D029742:Caenorhabditis elegans Proteins; D002352:Carrier Proteins; D029721:Drosophila Proteins; D005656:Fungal Proteins; D015801:Helminth Proteins; D051938:Munc18 Proteins; D009419:Nerve Tissue Proteins; D010750:Phosphoproteins; C086393:Rop protein, Drosophila; C497939:Stxbp1 protein, rat; D050827:Syntaxin 1; C082636:Unc-18 protein, C elegans; D033921:Vesicular Transport Proteins