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PMID: 8909535 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Amino acid permeases require COPII components and the ER resident membrane protein Shr3p for packaging into transport vesicles in vitro.

The Journal of cell biology ·Vol. 135 ·No. 3 ·1996-11-00 ·Pages 585-95

Kuehn MJ, Schekman R, Ljungdahl PO

Abstract

In S. cerevisiae lacking SHR3, amino acid permeases specifically accumulate in membranes of the endoplasmic reticulum (ER) and fail to be transported to the plasma membrane. We examined the requirements of transport of the permeases from the ER to the Golgi in vitro. Addition of soluble COPII components (Sec23/24p, Sec13/31p, and Sar1p) to yeast membrane preparations generated vesicles containing the general amino acid permease. Gap1p, and the histidine permease, Hip1p. Shr3p was required for the packaging of Gap1p and Hip1p but was not itself incorporated into transport vesicles. In contrast, the packaging of the plasma membrane ATPase, Pma1p, and the soluble yeast pheromone precursor, glycosylated pro alpha factor, was independent of Shr3p. In addition, we show that integral membrane and soluble cargo colocalize in transport vesicles, indicating that different types of cargo are not segregated at an early step in secretion. Our data suggest that specific ancillary proteins in the ER membrane recruit subsets of integral membrane protein cargo into COPII transport vesicles.

MeSH Terms
ATP-Binding Cassette Transporters Amino Acid Transport Systems Amino Acid Transport Systems, Basic Bacterial Proteins Biological Transport, Active COP-Coated Vesicles Endoplasmic Reticulum/metabolism Fungal Proteins/pharmacology GTP-Binding Proteins/physiology GTPase-Activating Proteins Golgi Apparatus/metabolism Intracellular Membranes/metabolism Mating Factor Membrane Proteins/pharmacology,physiology Membrane Transport Proteins/metabolism Microsomes/metabolism Monomeric GTP-Binding Proteins Nuclear Pore Complex Proteins Peptides/metabolism Protein Precursors/metabolism Proton-Translocating ATPases/metabolism Saccharomyces cerevisiae/enzymology Saccharomyces cerevisiae Proteins Spheroplasts/metabolism Vesicular Transport Proteins
Chemicals
ATP-Binding Cassette Transporters Amino Acid Transport Systems Amino Acid Transport Systems, Basic Bacterial Proteins Fungal Proteins GTPase-Activating Proteins Membrane Proteins Membrane Transport Proteins Nuclear Pore Complex Proteins PMA2 protein, S cerevisiae Peptides Protein Precursors SEC13 protein, S cerevisiae SEC23 protein, S cerevisiae SHR3 protein, S cerevisiae Saccharomyces cerevisiae Proteins Vesicular Transport Proteins Mating Factor histidine permease, Bacteria GTP-Binding Proteins PMA1 protein, S cerevisiae Proton-Translocating ATPases Monomeric GTP-Binding Proteins SAR1 protein, S cerevisiae
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Kuehn M J
Department of Molecular and Cell Biology, Howard Hughes Medical Institute, University of California, Berkeley 94720, USA.
Schekman R
Ljungdahl P O
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.135.3.585
Published
1996-11-00
Pages
585-95
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2121070
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.135.3.585 PMC Full Text View journal details
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