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PMID: 8089173 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Guanine nucleotide dissociation inhibitor is essential for Rab1 function in budding from the endoplasmic reticulum and transport through the Golgi stack.

The Journal of cell biology ·Vol. 126 ·No. 6 ·1994-09-00 ·Pages 1393-406

Peter F, Nuoffer C, Pind SN, Balch WE

Abstract

The small GTPase Rab1 is required for vesicular traffic from the ER to the cis-Golgi compartment, and for transport between the cis and medial compartments of the Golgi stack. In the present study, we examine the role of guanine nucleotide dissociation inhibitor (GDI) in regulating the function of Rab1 in the transport of vesicular stomatitis virus glycoprotein (VSV-G) in vitro. Incubation in the presence of excess GDI rapidly (t1/2 < 30 s) extracted Rab1 from membranes, inhibiting vesicle budding from the ER and sequential transport between the cis-, medial-, and trans-Golgi cisternae. These results demonstrate a direct role for GDI in the recycling of Rab proteins. Analysis of rat liver cytosol by gel filtration revealed that a major pool of Rab1 fractionates with a molecular mass of approximately 80 kD in the form of a GDI-Rab1 complex. When the GDI-Rab1 complex was depleted from cytosol by use of a Rab1-specific antibody, VSV-G failed to exit the ER. However, supplementation of depleted cytosol with a GDI-Rab1 complex prepared in vitro from recombinant forms of Rab1 and GDI efficiently restored export from the ER, and transport through the Golgi stack. These results provide evidence that a cytosolic GDI-Rab1 complex is required for the formation of non-clathrin-coated vesicles mediating transport through the secretory pathway.

MeSH Terms
Animals Biological Transport/physiology Cell Membrane Permeability Coatomer Protein Cytosol/chemistry Endoplasmic Reticulum/metabolism GTP Phosphohydrolases/analysis,physiology GTP-Binding Proteins/analysis,physiology Golgi Apparatus/metabolism Guanine Nucleotide Dissociation Inhibitors In Vitro Techniques Liver/chemistry Membrane Glycoproteins Microtubule-Associated Proteins/physiology Molecular Weight Rats Recombinant Proteins Vesicular stomatitis Indiana virus Viral Envelope Proteins/metabolism rab1 GTP-Binding Proteins
Chemicals
Coatomer Protein G protein, vesicular stomatitis virus GDP dissociation inhibitor 1 Guanine Nucleotide Dissociation Inhibitors Membrane Glycoproteins Microtubule-Associated Proteins Recombinant Proteins Viral Envelope Proteins GTP Phosphohydrolases GTP-Binding Proteins RAB1B protein, rat rab1 GTP-Binding Proteins
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Peter F
Department of Cell Biology, Scripps Research Institute, La Jolla, California 92037.
Nuoffer C
Pind S N
Balch W E
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.126.6.1393
Published
1994-09-00
Pages
1393-406
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2290957
Subset
IM
Grants
NIGMS NIH HHS · GM 42336 · United States
NCI NIH HHS · NCI CA58689 · United States
NIGMS NIH HHS · NCI GM 58689 · United States
External Links
PubMed source DOI: 10.1083/jcb.126.6.1393 PMC Full Text View journal details
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