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Cited by 61Cellular transport and secretionLipid Membrane Structure and BehaviorLipid metabolism and biosynthesis

A distinct tethering step is vital for vacuole membrane fusion

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Zick, Michael, Wickner, William T·2014·eLife
DOI 10.7554/elife.03251PMC PMC4200421
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Abstract

Past experiments with reconstituted proteoliposomes, employing assays that infer membrane fusion from fluorescent lipid dequenching, have suggested that vacuolar SNAREs alone suffice to catalyze membrane fusion in vitro. While we could replicate these results, we detected very little fusion with the more rigorous assay of lumenal compartment mixing. Exploring the discrepancies between lipid-dequenching and content-mixing assays, we surprisingly found that the disposition of the fluorescent lipids with respect to SNAREs had a striking effect. Without other proteins, the association of SNAREs in trans causes lipid dequenching that cannot be ascribed to fusion or hemifusion. Tethering of the SNARE-bearing proteoliposomes was required for efficient lumenal compartment mixing. While the physiological HOPS tethering complex caused a few-fold increase of trans-SNARE association, the rate of content mixing increased more than 100-fold. Thus tethering has a role in promoting membrane fusion that extends beyond simply increasing the amount of total trans-SNARE complex.

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  • Assays based on lipid mixing alone can produce misleading results since lipid mixing can occur without content mixing.

    “More importantly, assays based on lipid mixing alone can produce misleading results since lipid mixing can occur without content mixing86.”

    From Architecture of the synaptotagmin–SNARE machinery for neuronal exocytosis (2015) · cites this paper as [86]

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Cited by

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Authors
Zick, M., & Wickner, W. T.
Affiliations
Dartmouth College (US)
Corresponding author
  • Zick, Michael
  • Wickner, William T
ORCID
Wickner, William T
Venue
eLife
Publisher
eLife Sciences Publications, Ltd
Published
2014-09-25
Volume
3
Pages
e03251-e03251
ISSN
2050-084X (ISSN-L)
DOI
10.7554/elife.03251
PMC
PMC4200421
OpenAlex
W2123942529
Cited by
61
References
2
Citations by year
2026: 2 · 2024: 4 · 2023: 5 · 2022: 4 · 2021: 5 · 2020: 7 · 2019: 4 · 2018: 3 · 2017: 9 · 2016: 9 · 2015: 8 · 2014: 1
FWCI
4.48
Type
journal article
Language
en
Open access
Licence
Locations
  • eLife (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
National Institutes of Health (R01 GM23377-40)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Lipid Membrane Structure and Behavior
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Biochemistry › Lipid metabolism and biosynthesis
Keywords
SNARE proteins, tethering, HOPS complex, proteoliposomes, trans-SNARE complex, hemifusion
MeSH terms
Fluorescent Dyes, Inositol Phosphates, Lipids, Membrane Fusion, Protein Binding, Vacuoles, Biomarkers, Multiprotein Complexes, Soluble N-Ethylmaleimide-Sensitive Factor Attachment Proteins

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