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PMID: 1894692 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

PAS3, a Saccharomyces cerevisiae gene encoding a peroxisomal integral membrane protein essential for peroxisome biogenesis.

The Journal of cell biology ·Vol. 114 ·No. 6 ·1991-09-00 ·Pages 1167-78

Höhfeld J, Veenhuis M, Kunau WH

Abstract

Saccharomyces cerevisiae pas3-mutants are described which conform the pas-phenotype recently reported for the peroxisomal assembly mutants pas1-1 and pas2 (Erdmann, R., M. Veenhuis, D. Mertens, and W.-H Kunau, 1989, Proc. Natl. Acad. Sci. USA. 86:5419-5423). The isolation of pas3-mutants enabled us to clone the PAS3 gene by functional complementation. DNA sequence analysis revealed a 50.6-kD protein with at least one domain of sufficient length and hydrophobicity to span a lipid bilayer. To verify these predictions antibodies were raised against a truncated portion of the PAS3 coding region overexpressed in E. coli. Pas3p was identified as a 48 kD peroxisomal integral membrane protein. It is shown that a lack of this protein causes the peroxisome-deficient phenotype and the cytosolic mislocalization of peroxisomal matrix enzymes. Based on protease digestion experiments Pas3p is discussed to be anchored in the peroxisomal membrane by its amino-terminus while the bulk of the molecule is exposed to the cytosol. These findings are consistent with the possibility that Pas3p is one component of the peroxisomal import machinery.

MeSH Terms
ATP-Binding Cassette Transporters Amino Acid Sequence Base Sequence Cloning, Molecular Escherichia coli/genetics Fungal Proteins/genetics,isolation & purification Genes, Fungal Genetic Complementation Test Genotype Immunoblotting Membrane Proteins/genetics,isolation & purification Microbodies/physiology,ultrastructure Microscopy, Electron Models, Structural Molecular Sequence Data Molecular Weight Peroxins Phenotype Protein Conformation Recombinant Proteins/isolation & purification Restriction Mapping Saccharomyces cerevisiae/genetics,physiology,ultrastructure Saccharomyces cerevisiae Proteins
Chemicals
ATP-Binding Cassette Transporters Fungal Proteins Membrane Proteins PEX3 protein, S cerevisiae Peroxins Recombinant Proteins SPF1 protein, S cerevisiae Saccharomyces cerevisiae Proteins
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Höhfeld J
Abteilung für Zellbiochemie, Ruhr-Universität Bochum, Federal Republic of Germany.
Veenhuis M
Kunau W H
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.114.6.1167
Published
1991-09-00
Pages
1167-78
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2289127
Subset
IM
Databases
GENBANK
M60691, M60692, M60693, M60694, M60695, M60696, M60697, S55275, X56699, X57971, X58407
External Links
PubMed source DOI: 10.1083/jcb.114.6.1167 PMC Full Text View journal details
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