The peroxin pex3p initiates membrane assembly in peroxisome biogenesis

Rat cDNA encoding a 372-amino-acid peroxin was isolated, primarily by functional complementation screening, using a peroxisome-deficient Chinese hamster ovary cell mutant, ZPG208, of complementation group 17. The deduced primary sequence showed approximately 25% amino acid identity with the yeast Pe...

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Bibliographic Details
Published in:Molecular biology of the cell 2000-06, Vol.11 (6), p.2085-2102
Main Authors: Ghaedi, K, Tamura, S, Okumoto, K, Matsuzono, Y, Fujiki, Y
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Animals
ATP-Binding Cassette Transporters
Base Sequence
Carrier Proteins - genetics
Carrier Proteins - metabolism
CHO Cells
Cloning, Molecular
Cricetinae
DNA, Complementary
Fungal Proteins
Humans
Intracellular Fluid - metabolism
Intracellular Membranes - metabolism
Kinetics
Membrane Proteins - genetics
Membrane Proteins - metabolism
Molecular Sequence Data
Peroxins
Peroxisomes - metabolism
Peroxisomes - physiology
Rats
Saccharomyces cerevisiae Proteins
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Summary:Rat cDNA encoding a 372-amino-acid peroxin was isolated, primarily by functional complementation screening, using a peroxisome-deficient Chinese hamster ovary cell mutant, ZPG208, of complementation group 17. The deduced primary sequence showed approximately 25% amino acid identity with the yeast Pex3p, thereby we termed this cDNA rat PEX3 (RnPEX3). Human and Chinese hamster Pex3p showed 96 and 94% identity to rat Pex3p and had 373 amino acids. Pex3p was characterized as an integral membrane protein of peroxisomes, exposing its N- and C-terminal parts to the cytosol. A homozygous, inactivating missense mutation, G to A at position413, in a codon (GGA) for Gly(138) and resulting in a codon (GAA) for Glu was the genetic cause of peroxisome deficiency of complementation group 17 ZPG208. The peroxisome-restoring activity apparently required the full length of Pex3p, whereas its N-terminal part from residues 1 to 40 was sufficient to target a fusion protein to peroxisomes. We also demonstrated that Pex3p binds the farnesylated peroxisomal membrane protein Pex19p. Moreover, upon expression of PEX3 in ZPG208, peroxisomal membrane vesicles were assembled before the import of soluble proteins such as PTS2-tagged green fluorescent protein. Thus, Pex3p assembles membrane vesicles before the matrix proteins are translocated.
ISSN:1059-1524
1939-4586
DOI:10.1091/mbc.11.6.2085