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PMID: 11739408 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

The Tlg SNARE complex is required for TGN homotypic fusion.

The Journal of cell biology ·Vol. 155 ·No. 6 ·2001-12-10 ·Pages 969-78

Brickner JH, Blanchette JM, Sipos G, Fuller RS

Abstract

Using a new assay for membrane fusion between late Golgi/endosomal compartments, we have reconstituted a rapid, robust homotypic fusion reaction between membranes containing Kex2p and Ste13p, two enzymes resident in the yeast trans-Golgi network (TGN). Fusion was temperature, ATP, and cytosol dependent. It was inhibited by dilution, Ca+2 chelation, N-ethylmaleimide, and detergent. Coimmunoisolation confirmed that the reaction resulted in cointegration of the two enzymes into the same bilayer. Antibody inhibition experiments coupled with antigen competition indicated a requirement for soluble NSF attachment protein receptor (SNARE) proteins Tlg1p, Tlg2p, and Vti1p in this reaction. Membrane fusion also required the rab protein Vps21p. Vps21p was sufficient if present on either the Kex2p or Ste13p membranes alone, indicative of an inherent symmetry in the reaction. These results identify roles for a Tlg SNARE complex composed of Tlg1p, Tlg2p, Vti1p, and the rab Vps21p in this previously uncharacterized homotypic TGN fusion reaction.

MeSH Terms
Carrier Proteins/metabolism Cell-Free System Fucosyltransferases/metabolism Membrane Fusion/physiology Membrane Proteins/genetics,metabolism Membrane Transport Proteins Qa-SNARE Proteins Qb-SNARE Proteins SNARE Proteins Saccharomyces cerevisiae Proteins Vesicular Transport Proteins Yeasts rab GTP-Binding Proteins/metabolism rab5 GTP-Binding Proteins/metabolism trans-Golgi Network/metabolism
Chemicals
Carrier Proteins Membrane Proteins Membrane Transport Proteins Qa-SNARE Proteins Qb-SNARE Proteins SNARE Proteins Saccharomyces cerevisiae Proteins TLG2 protein, S cerevisiae VTI1 protein, S cerevisiae Vesicular Transport Proteins Fucosyltransferases galactoside 2-alpha-L-fucosyltransferase rab GTP-Binding Proteins rab5 GTP-Binding Proteins
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Brickner J H
Department of Biological Chemistry, University of Michigan, Ann Arbor, MI 48109, USA.
Blanchette J M
Sipos G
Fuller R S
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.200104093
Published
2001-12-10
Epub
2001-00-10
Pages
969-78
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2150899
Subset
IM
Grants
NIGMS NIH HHS · R01 GM050915 · United States
NIGMS NIH HHS · T32 GM007544 · United States
NIGMS NIH HHS · GM07544 · United States
NIGMS NIH HHS · GM50915 · United States
External Links
PubMed source DOI: 10.1083/jcb.200104093 PMC Full Text View journal details
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