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PMID: 9199167 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

The yeast v-SNARE Vti1p mediates two vesicle transport pathways through interactions with the t-SNAREs Sed5p and Pep12p.

The Journal of cell biology ·Vol. 137 ·No. 7 ·1997-06-30 ·Pages 1511-24

von Mollard GF, Nothwehr SF, Stevens TH

Abstract

Membrane traffic in eukaryotic cells requires that specific v-SNAREs on transport vesicles interact with specific t-SNAREs on target membranes. We identified a novel Saccharomyces cerevisiae v-SNARE (Vti1p) encoded by the essential gene, VTI1. Vti1p interacts with the prevacuolar t-SNARE Pep12p to direct Golgi to prevacuolar traffic. vti1-1 mutant cells missorted and secreted the soluble vacuolar hydrolase carboxypeptidase Y (CPY) rapidly and reversibly when vti1-1 cells were shifted to the restrictive temperature. However, overexpression of Pep12p suppressed the CPY secretion defect exhibited by vti1-1 cells at 36 degrees C. Characterization of a second vti1 mutant, vti1-11, revealed that Vti1p also plays a role in membrane traffic at a cis-Golgi stage. vti1-11 mutant cells displayed a growth defect and accumulated the ER and early Golgi forms of both CPY and the secreted protein invertase at the nonpermissive temperature. Overexpression of the yeast cis-Golgi t-SNARE Sed5p suppressed the accumulation of the ER form of CPY but did not lead to CPY transport to the vacuole in vti1-11 cells. Overexpression of Sed5p allowed growth in the absence of Vti1p. In vitro binding and coimmunoprecipitation studies revealed that Vti1p interacts directly with the two t-SNAREs, Sed5p and Pep12p. These data suggest that Vti1p plays a role in cis-Golgi membrane traffic, which is essential for yeast viability, and a nonessential role in the fusion of Golgi-derived vesicles with the prevacuolar compartment. Therefore, a single v-SNARE can interact functionally with two different t-SNAREs in directing membrane traffic in yeast.

MeSH Terms
Biological Transport/genetics Carrier Proteins/genetics,metabolism Cytoplasmic Granules/genetics,metabolism Fungal Proteins/genetics,metabolism Gene Expression Regulation, Fungal Membrane Proteins/genetics,metabolism Qa-SNARE Proteins Qb-SNARE Proteins Saccharomyces cerevisiae/genetics,metabolism,ultrastructure Saccharomyces cerevisiae Proteins Vesicular Transport Proteins
Chemicals
Carrier Proteins Fungal Proteins Membrane Proteins PEP12 protein, S cerevisiae Qa-SNARE Proteins Qb-SNARE Proteins Saccharomyces cerevisiae Proteins Sed5 protein, S cerevisiae VTI1 protein, S cerevisiae Vesicular Transport Proteins
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
von Mollard G F
Institute of Molecular Biology, University of Oregon, Eugene, Oregon 97403-1229, USA.
Nothwehr S F
Stevens T H
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.137.7.1511
Published
1997-06-30
Pages
1511-24
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2137825
Subset
IM
Grants
NIGMS NIH HHS · GM32448 · United States
Databases
GENBANK
AF006074
External Links
PubMed source DOI: 10.1083/jcb.137.7.1511 PMC Full Text View journal details
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