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PMID: 8019009 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Cytosolic ARFs are required for vesicle formation but not for cell-free intra-Golgi transport: evidence for coated vesicle-independent transport.

Molecular biology of the cell ·Vol. 5 ·No. 2 ·1994-02-00 ·Pages 237-52

Taylor TC, Kanstein M, Weidman P, Melançon P

Abstract

We investigated the role of ADP-ribosylation factors (ARFs) in Golgi function using biochemical and morphological cell-free assays. An ARF-free cytosol produced from soluble Chinese hamster ovary (CHO) extracts supports intra-Golgi transport by a mechanism that is biochemically indistinguishable from control transport reactions: ARF-free transport reactions are NSF-dependent, remain sensitive to the donor Golgi-specific inhibitor ilimaquinone, and exhibit kinetics that are identical to that of control reactions containing ARFs. In contrast, ARF-free cytosol does not support the formation of coated vesicles on Golgi cisternae. However, vesicle formation is reconstituted upon the addition of ARF1. These data suggest that neither soluble ARFs nor coated vesicle formation are essential for transport. We conclude that cell-free intra-Golgi transport proceeds via a coated vesicle-independent mechanism regardless of vesicle formation on Golgi cisternae.

MeSH Terms
ADP-Ribosylation Factor 1 ADP-Ribosylation Factors Animals Biological Transport/drug effects CHO Cells Cell-Free System Coated Pits, Cell-Membrane/metabolism Cricetinae Cricetulus Cytosol/metabolism Ethylmaleimide/pharmacology GTP-Binding Proteins/metabolism Glycosylation Golgi Apparatus/metabolism,ultrastructure Intracellular Membranes/metabolism,ultrastructure Kinetics Quinones/pharmacology
Chemicals
Quinones illimaquinone GTP-Binding Proteins ADP-Ribosylation Factor 1 ADP-Ribosylation Factors Ethylmaleimide
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Taylor T C
Department of Chemistry and Biochemistry, University of Colorado, Boulder 80303-0215.
Kanstein M
Weidman P
Melançon P
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Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.5.2.237
Published
1994-02-00
Pages
237-52
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC301029
Subset
IM
Grants
NIGMS NIH HHS · 5R01GM-29647 · United States
NIGMS NIH HHS · 5R01GM-43378 · United States
External Links
PubMed source DOI: 10.1091/mbc.5.2.237 PMC Full Text View journal details
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