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PMID: 9647644 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

The SNARE machinery is involved in apical plasma membrane trafficking in MDCK cells.

The Journal of cell biology ·Vol. 141 ·No. 7 ·1998-06-29 ·Pages 1503-13

Low SH, Chapin SJ, Wimmer C, Whiteheart SW, Kömüves LG, Mostov KE, Weimbs T

Abstract

We have investigated the controversial involvement of components of the SNARE (soluble N-ethyl maleimide-sensitive factor [NSF] attachment protein [SNAP] receptor) machinery in membrane traffic to the apical plasma membrane of polarized epithelial (MDCK) cells. Overexpression of syntaxin 3, but not of syntaxins 2 or 4, caused an inhibition of TGN to apical transport and apical recycling, and leads to an accumulation of small vesicles underneath the apical plasma membrane. All other tested transport steps were unaffected by syntaxin 3 overexpression. Botulinum neurotoxin E, which cleaves SNAP-23, and antibodies against alpha-SNAP inhibit both TGN to apical and basolateral transport in a reconstituted in vitro system. In contrast, we find no evidence for an involvement of N-ethyl maleimide-sensitive factor in TGN to apical transport, whereas basolateral transport is NSF-dependent. We conclude that syntaxin 3, SNAP-23, and alpha-SNAP are involved in apical membrane fusion. These results demonstrate that vesicle fusion with the apical plasma membrane does not use a mechanism that is entirely unrelated to other cellular membrane fusion events, but uses isoforms of components of the SNARE machinery, which suggests that they play a role in providing specificity to polarized membrane traffic.

MeSH Terms
Animals Biological Transport Carrier Proteins/metabolism Cell Line Cell Membrane/metabolism Cell Membrane Permeability Cell Polarity Coated Vesicles/metabolism Dogs Endocytosis Immunoglobulin A/metabolism Membrane Proteins/genetics,metabolism N-Ethylmaleimide-Sensitive Proteins Qa-SNARE Proteins Qb-SNARE Proteins Qc-SNARE Proteins SNARE Proteins Soluble N-Ethylmaleimide-Sensitive Factor Attachment Proteins Vesicular Transport Proteins
Chemicals
Carrier Proteins Immunoglobulin A Membrane Proteins Qa-SNARE Proteins Qb-SNARE Proteins Qc-SNARE Proteins SNAP23 protein, human SNARE Proteins Soluble N-Ethylmaleimide-Sensitive Factor Attachment Proteins Vesicular Transport Proteins N-Ethylmaleimide-Sensitive Proteins
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Low S H
Department of Anatomy, Biochemistry and Biophysics, Cardiovascular Research Institute, University of California, San Francisco, California 94143-0452, USA.
Chapin S J
Wimmer C
Whiteheart S W
Kömüves L G
Mostov K E
Weimbs T
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.141.7.1503
Published
1998-06-29
Pages
1503-13
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2133007
Subset
IM
Grants
NIAID NIH HHS · AI39161 · United States
NHLBI NIH HHS · T32HL07731 · United States
NIAID NIH HHS · R01 AI25144 · United States
External Links
PubMed source DOI: 10.1083/jcb.141.7.1503 PMC Full Text View journal details
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