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Amino acid permeases require COPII components and the ER resident membrane protein Shr3p for packaging into transport vesicles in vitro.
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.
Type
Journal articleSubject
ATP-Binding Cassette TransportersAmino Acid Transport Systems
Amino Acid Transport Systems, Basic
Bacterial Proteins
Biological Transport, Active
COP-Coated Vesicles
Endoplasmic Reticulum
Fungal Proteins
GTP-Binding Proteins
GTPase-Activating Proteins
Golgi Apparatus
Intracellular Membranes
Mating Factor
Membrane Proteins
Membrane Transport Proteins
Microsomes
Monomeric GTP-Binding Proteins
Nuclear Pore Complex Proteins
Peptides
Protein Precursors
Proton-Translocating ATPases
Saccharomyces cerevisiae
Saccharomyces cerevisiae Proteins
Spheroplasts
Vesicular Transport Proteins
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Show full item recordScholars@Duke
Margarethe Joanna Kuehn
Associate Professor of Biochemistry
Enterotoxigenic E. coli (ETEC) causes traveler's diarrhea and infant mortality in
underdeveloped countries, and Pseudomonas aeruginosa is an opportunistic pathogen
for immunocompromised patients. Like all gram negative bacteria studied to date, ETEC
and P. aeruginosa produce small outer membrane vesicles that can serve as delivery
"bombs" to host tissues. Vesicles contain a subset of outer membrane and soluble periplasmic
proteins and lipids. In tissues and sera of infected hosts, vesicles have

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