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PMID: 14734531 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Trans-SNARE interactions elicit Ca2+ efflux from the yeast vacuole lumen.

The Journal of cell biology ·Vol. 164 ·No. 2 ·2004-01-19 ·Pages 195-206

Merz AJ, Wickner WT

Abstract

Ca2+ transients trigger many SNARE-dependent membrane fusion events. The homotypic fusion of yeast vacuoles occurs after a release of lumenal Ca2+. Here, we show that trans-SNARE interactions promote the release of Ca2+ from the vacuole lumen. Ypt7p-GTP, the Sec1p/Munc18-protein Vps33p, and Rho GTPases, all of which function during docking, are required for Ca2+ release. Inhibitors of SNARE function prevent Ca2+ release. Recombinant Vam7p, a soluble Q-SNARE, stimulates Ca2+ release. Vacuoles lacking either of two complementary SNAREs, Vam3p or Nyv1p, fail to release Ca2+ upon tethering. Mixing these two vacuole populations together allows Vam3p and Nyv1p to interact in trans and rescues Ca2+ release. Sec17/18p promote sustained Ca2+ release by recycling SNAREs (and perhaps other limiting factors), but are not required at the release step itself. We conclude that trans-SNARE assembly events during docking promote Ca2+ release from the vacuole lumen.

MeSH Terms
Calcium/metabolism Kinetics Membrane Fusion Membrane Proteins/metabolism Nerve Tissue Proteins/metabolism Recombinant Proteins/metabolism SNARE Proteins Saccharomyces cerevisiae/physiology Saccharomyces cerevisiae Proteins/metabolism Synaptosomal-Associated Protein 25 Vacuoles/physiology Vesicular Transport Proteins/metabolism
Chemicals
Membrane Proteins Nerve Tissue Proteins Recombinant Proteins SNARE Proteins Saccharomyces cerevisiae Proteins Synaptosomal-Associated Protein 25 Vesicular Transport Proteins Calcium
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Merz Alexey J
Dept. of Biochemistry, 7200 Vail Bldg., Dartmouth Medical School, Hanover, NH 03755-3844, USA.
Wickner William T
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.200310105
Published
2004-01-19
Pages
195-206
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2172329
Subset
IM
Grants
NIAMS NIH HHS · T32 AR007576 · United States
NIAMS NIH HHS · AR07576-09 · United States
External Links
PubMed source DOI: 10.1083/jcb.200310105 PMC Full Text View journal details
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