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PMID: 10601335 Published · ppublish English Journal Article

ER to Golgi transport: Requirement for p115 at a pre-Golgi VTC stage.

The Journal of cell biology ·Vol. 147 ·No. 6 ·1999-12-13 ·Pages 1205-22

Alvarez C, Fujita H, Hubbard A, Sztul E

Abstract

The membrane transport factor p115 functions in the secretory pathway of mammalian cells. Using biochemical and morphological approaches, we show that p115 participates in the assembly and maintenance of normal Golgi structure and is required for ER to Golgi traffic at a pre-Golgi stage. Injection of antibodies against p115 into intact WIF-B cells caused Golgi disruption and inhibited Golgi complex reassembly after BFA treatment and wash-out. Addition of anti-p115 antibodies or depletion of p115 from a VSVtsO45 based semi-intact cell transport assay inhibited transport. The inhibition occurred after VSV glycoprotein (VSV-G) exit from the ER but before its delivery to the Golgi complex, and resulted in VSV-G protein accumulating in peripheral vesicular tubular clusters (VTCs). The p115-requiring step of transport followed the rab1-requiring step and preceded the Ca(2+)-requiring step. Unexpectedly, mannosidase I redistributed from the Golgi complex to colocalize with VSV-G protein arrested in pre-Golgi VTCs by p115 depletion. Redistribution of mannosidase I was also observed in cells incubated at 15 degrees C. Our data show that p115 is essential for the translocation of pre-Golgi VTCs from peripheral sites to the Golgi stack. This defines a previously uncharacterized function for p115 at the VTC stage of ER to Golgi traffic.

MeSH Terms
Animals Antibodies, Monoclonal/immunology,pharmacology Biological Transport/drug effects Brefeldin A/pharmacology Calcium/physiology Carrier Proteins/antagonists & inhibitors,chemistry,immunology,metabolism Cell Line Endoplasmic Reticulum/drug effects,metabolism Fluorescent Antibody Technique Glycosylation Golgi Apparatus/drug effects,enzymology,metabolism Golgi Matrix Proteins Mannosidases/metabolism Membrane Glycoproteins Membrane Proteins/antagonists & inhibitors,chemistry,immunology,metabolism N-Acetylglucosaminyltransferases/metabolism Organelles/metabolism Rats Temperature Time Factors Vesicular Transport Proteins Viral Envelope Proteins/metabolism rab1 GTP-Binding Proteins/immunology,physiology
Chemicals
Antibodies, Monoclonal Carrier Proteins G protein, vesicular stomatitis virus Golgi Matrix Proteins Membrane Glycoproteins Membrane Proteins Vesicular Transport Proteins Viral Envelope Proteins vesicular transport factor p115 Brefeldin A N-Acetylglucosaminyltransferases Mannosidases mannosyl-oligosaccharide 1,2-alpha-mannosidase mannosyl-oligosaccharide 1,3 - 1,6-alpha-mannosidase rab1 GTP-Binding Proteins Calcium
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Alvarez C
Department of Cell Biology, University of Alabama at Birmingham, Birmingham, Alabama 35294, USA.
Fujita H
Hubbard A
Sztul E
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.147.6.1205
Published
1999-12-13
Pages
1205-22
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2168100
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.147.6.1205 PMC Full Text View journal details
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