Sec34p, a protein required for vesicle tethering to the yeast Golgi apparatus, is in a complex with Sec35p
| Autor: | Stephanie K. Sapperstein, Vladimir Lupashin, Xiaochun Cao, Elbert C. Chiang, M. Gerard Waters, Susan M. VanRheenen, Charles Barlowe |
|---|---|
| Rok vydání: | 1999 |
| Předmět: |
vesicle tethering
Saccharomyces cerevisiae Proteins Genotype Mutant Molecular Sequence Data Restriction Mapping Vesicular Transport Proteins Golgi Apparatus Saccharomyces cerevisiae Biology Cell Fractionation Vesicle tethering Fungal Proteins 03 medical and health sciences symbols.namesake Sec34p 0302 clinical medicine Amino Acid Sequence Cloning Molecular Secretory pathway 030304 developmental biology 0303 health sciences Binding Sites Conserved oligomeric Golgi complex USO1 Membrane Proteins Cell Biology Golgi apparatus Recombinant Proteins Cell biology Transport protein secretory pathway Sec35p Adaptor Proteins Vesicular Transport symbols Original Article Rab Carrier Proteins Rud3p 030217 neurology & neurosurgery Gene Deletion Plasmids Protein Binding |
| Zdroj: | The Journal of Cell Biology |
| ISSN: | 0021-9525 |
| Popis: | A screen for mutants of Saccharomyces cerevisiae secretory pathway components previously yielded sec34, a mutant that accumulates numerous vesicles and fails to transport proteins from the ER to the Golgi complex at the restrictive temperature (Wuestehube, L.J., R. Duden, A. Eun, S. Hamamoto, P. Korn, R. Ram, and R. Schekman. 1996. Genetics. 142:393–406). We find that SEC34 encodes a novel protein of 93-kD, peripherally associated with membranes. The temperature-sensitive phenotype of sec34-2 is suppressed by the rab GTPase Ypt1p that functions early in the secretory pathway, or by the dominant form of the ER to Golgi complex target-SNARE (soluble N-ethylmaleimide sensitive fusion protein attachment protein receptor)–associated protein Sly1p, Sly1-20p. Weaker suppression is evident upon overexpression of genes encoding the vesicle tethering factor Uso1p or the vesicle-SNAREs Sec22p, Bet1p, or Ykt6p. This genetic suppression profile is similar to that of sec35-1, a mutant allele of a gene encoding an ER to Golgi vesicle tethering factor and, like Sec35p, Sec34p is required in vitro for vesicle tethering. sec34-2 and sec35-1 display a synthetic lethal interaction, a genetic result explained by the finding that Sec34p and Sec35p can interact by two-hybrid analysis. Fractionation of yeast cytosol indicates that Sec34p and Sec35p exist in an ∼750-kD protein complex. Finally, we describe RUD3, a novel gene identified through a genetic screen for multicopy suppressors of a mutation in USO1, which suppresses the sec34-2 mutation as well. |
| Databáze: | OpenAIRE |
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