Abstract
Many of the molecules necessary for neurotransmission are homologous to proteins involved in the Golgi-to-plasma membrane stage of the yeast secretory pathway. Of 15 genes known to be essential for the later stages of vesicle trafficking in yeast, 7 have no identified mammalian homologs. These include the yeast SEC6, SEC8, and SEC15 genes, whose products are constituents of a 19.5S particle that interacts with the GTP-binding protein Sec4p. Here we report the sequences of rSec6 and rSec8, rat homologs of Sec6p and Sec8p. The rSec6 cDNA is predicted to encode an 87-kDa protein with 22% amino acid identity to Sec6p, and the rSec8 cDNA is predicted to encode a 110-kDa protein which is 20% identical to Sec8p. Northern blot analysis indicates that rSec6 and rSec8 are expressed in similar tissues. Immunodetection reveals that rSec8 is part of a soluble 17S particle in brain. COS cell cotransfection studies demonstrate that rSec8 colocalizes with the GTP-binding protein Rab3a and syntaxin 1a, two proteins involved in synaptic vesicle docking and fusion at the presynaptic terminal. These data suggest that rSec8 is a component of a high molecular weight complex which may participate in the regulation of vesicle docking and fusion in brain.
MeSH Terms
Amino Acid Sequence
Animals
Antigens, Surface/metabolism
Base Sequence
Biological Transport
Blotting, Northern
Blotting, Western
Brain
Carrier Proteins
Cloning, Molecular
Fluorescent Antibody Technique
GTP-Binding Proteins/metabolism
Gene Library
Macromolecular Substances
Membrane Proteins
Models, Biological
Molecular Sequence Data
Nerve Tissue Proteins/genetics,isolation & purification,metabolism
Protein Binding
RNA, Messenger/analysis
Rats
Recombinant Proteins/metabolism
Sequence Analysis, DNA
Sequence Homology, Amino Acid
Synaptic Transmission
Syntaxin 1
Yeasts/genetics
rab3 GTP-Binding Proteins
Chemicals
Antigens, Surface
Carrier Proteins
Exoc3 protein, rat
Exoc4 protein, rat
Macromolecular Substances
Membrane Proteins
Nerve Tissue Proteins
RNA, Messenger
Recombinant Proteins
Stx1a protein, rat
Syntaxin 1
GTP-Binding Proteins
rab3 GTP-Binding Proteins
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Ting A E
Department of Molecular and Cellular Physiology, Howard Hughes Medical Institute, Stanford University Medical Center, CA 94305-5428, USA.
Hazuka C D
Hsu S C
Kirk M D
Bean A J
Scheller R H
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