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PMID: 8408203 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Rab8, a small GTPase involved in vesicular traffic between the TGN and the basolateral plasma membrane.

The Journal of cell biology ·Vol. 123 ·No. 1 ·1993-10-00 ·Pages 35-45

Huber LA, Pimplikar S, Parton RG, Virta H, Zerial M, Simons K

Abstract

Small GTP-binding proteins of the rab family have been implicated as regulators of membrane traffic along the biosynthetic and endocytic pathways in eukaryotic cells. We have investigated the localization and function of rab8, closely related to the yeast YPT1/SEC4 gene products. Confocal immunofluorescence microscopy and immunoelectron microscopy on filter-grown MDCK cells demonstrated that, rab8 was localized to the Golgi region, vesicular structures, and to the basolateral plasma membrane. Two-dimensional gel electrophoresis showed that rab8p was highly enriched in immuno-isolated basolateral vesicles carrying vesicular stomatitis virus-glycoprotein (VSV-G) but was absent from vesicles transporting the hemagglutinin protein (HA) of influenza virus to the apical cell surface. Using a cytosol dependent in vitro transport assay in permeabilized MDCK cells we studied the functional role of rab8 in biosynthetic membrane traffic. Transport of VSV-G from the TGN to the basolateral plasma membrane was found to be significantly inhibited by a peptide derived from the hypervariable COOH-terminal region of rab8, while transport of the influenza HA from the TGN to the apical surface and ER to Golgi transport were unaffected. We conclude that rab8 plays a role in membrane traffic from the TGN to the basolateral plasma membrane in MDCK cells.

MeSH Terms
ADP-Ribosylation Factors Animals Base Sequence Biological Transport Cell Membrane/metabolism Cell Polarity Cells, Cultured Coatomer Protein DNA-Directed RNA Polymerases/genetics Fluorescent Antibody Technique GTP-Binding Proteins/genetics,isolation & purification,metabolism Golgi Apparatus/metabolism Hemagglutinin Glycoproteins, Influenza Virus Hemagglutinins, Viral/metabolism Humans Membrane Glycoproteins Microtubule-Associated Proteins/isolation & purification Molecular Sequence Data RNA, Messenger/genetics Recombinant Proteins/metabolism Transfection Vaccinia virus/genetics Viral Envelope Proteins/metabolism Viral Proteins rab GTP-Binding Proteins
Chemicals
Coatomer Protein G protein, vesicular stomatitis virus Hemagglutinin Glycoproteins, Influenza Virus Hemagglutinins, Viral Membrane Glycoproteins Microtubule-Associated Proteins RNA, Messenger Recombinant Proteins Viral Envelope Proteins Viral Proteins bacteriophage T7 RNA polymerase DNA-Directed RNA Polymerases GTP-Binding Proteins RAB8A protein, human ADP-Ribosylation Factors rab GTP-Binding Proteins
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Huber L A
Cell Biology Programme, European Molecular Biology Laboratory, Heidelberg, Germany.
Pimplikar S
Parton R G
Virta H
Zerial M
Simons K
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.123.1.35
Published
1993-10-00
Pages
35-45
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2119815
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.123.1.35 PMC Full Text View journal details
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