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PMID: 11038190 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Geranylgeranylated SNAREs are dominant inhibitors of membrane fusion.

The Journal of cell biology ·Vol. 151 ·No. 2 ·2000-10-16 ·Pages 453-66

Grote E, Baba M, Ohsumi Y, Novick PJ

Abstract

Exocytosis in yeast requires the assembly of the secretory vesicle soluble N-ethylmaleimide-sensitive factor attachment protein receptor (v-SNARE) Sncp and the plasma membrane t-SNAREs Ssop and Sec9p into a SNARE complex. High-level expression of mutant Snc1 or Sso2 proteins that have a COOH-terminal geranylgeranylation signal instead of a transmembrane domain inhibits exocytosis at a stage after vesicle docking. The mutant SNARE proteins are membrane associated, correctly targeted, assemble into SNARE complexes, and do not interfere with the incorporation of wild-type SNARE proteins into complexes. Mutant SNARE complexes recruit GFP-Sec1p to sites of exocytosis and can be disassembled by the Sec18p ATPase. Heterotrimeric SNARE complexes assembled from both wild-type and mutant SNAREs are present in heterogeneous higher-order complexes containing Sec1p that sediment at greater than 20S. Based on a structural analogy between geranylgeranylated SNAREs and the GPI-HA mutant influenza virus fusion protein, we propose that the mutant SNAREs are fusion proteins unable to catalyze fusion of the distal leaflets of the secretory vesicle and plasma membrane. In support of this model, the inverted cone-shaped lipid lysophosphatidylcholine rescues secretion from SNARE mutant cells.

MeSH Terms
Cell Membrane/metabolism,ultrastructure Diterpenes/metabolism Exocytosis/drug effects Fungal Proteins/genetics,metabolism Lipoproteins/pharmacology Membrane Fusion/drug effects Membrane Proteins/genetics,metabolism Models, Biological Mutation Protein Prenylation Protein Structure, Tertiary Qa-SNARE Proteins R-SNARE Proteins SNARE Proteins Saccharomyces cerevisiae Proteins Secretory Vesicles/metabolism,ultrastructure Vesicular Transport Proteins Yeasts/physiology
Chemicals
Diterpenes Fungal Proteins Lipoproteins Membrane Proteins Qa-SNARE Proteins R-SNARE Proteins SNARE Proteins SNC1 protein, S cerevisiae SSO2 protein, S cerevisiae Saccharomyces cerevisiae Proteins Vesicular Transport Proteins geranylgeraniol
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Grote E
Department of Cell Biology, Yale University School of Medicine, New Haven, Connecticut 06520, USA.
Baba M
Ohsumi Y
Novick P J
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.151.2.453
Published
2000-10-16
Pages
453-66
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2192637
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.151.2.453 PMC Full Text View journal details
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