title: PEB1 (PAS7) in Saccharomyces cerevisiae encodes a hydrophilic, intra-peroxisomal protein that is a member of the WD repeat family and is essential for the import of thiolase into peroxisomes.
abstract: We have previously described mutant S. cerevisiae that are defective in peroxisome biogenesis (peb mutants) (Zhang, J. W., Y. Han, and P. B. Lazarow. 1993. J. Cell Biol. 123:1133-1147.). In some mutants, peroxisomes are undetectable. Other mutants contain normal-looking peroxisomes but fail to package subsets of peroxisomal proteins into the organelle (Zhang, J. W., C. Luckey, and P. B. Lazarow. 1993. Mol. Biol. Cell. 4:1351-1359.). In peb1 (pas7) cells, for example, the peroxisomes contain proteins that are targeted by COOH-terminal tripeptides and contain acyl-CoA oxidase (which is probably targeted by internal oligopeptides), but fail to import thiolase (which is targeted by an NH(2)-terminal 16-amino acid sequence). These and other data suggest that there are three branches in the pathway for the import of proteins into peroxisomes, each of which contains a receptor for one type of peroxisomal topogenic information. Here, we report the cloning and characterization of the PEB1 gene, that encodes a 42,320-Da hydrophilic protein with no predicted transmembrane segment. The protein contains six WD repeats, a motif which has been found in 27 proteins involved in diverse cellular functions. The PEB1 gene product was tagged with the hemagglutinin epitope and found to rescue thiolase import in the peb1 null mutant. The epitope-tagged protein was shown to be inside of peroxisomes by immunofluorescence, digitonin permeabilization, equilibrium density centrifugation, immunoelectron microscopy, and proteinase K protection studies. The PEB1 gene product does not cleave the thiolase-targeting sequence. It may function to draw thiolase into peroxisomes.
authors.affiliation: Department of Cell Biology and Anatomy, Mount Sinai School of Medicine, New York 10029.
authors.forename: J W
authors.initials: JW
authors.lastname: Zhang
authors.forename: P B
authors.initials: PB
authors.lastname: Lazarow
mesh_terms: D000101:Acetyl-CoA C-Acetyltransferase / Q000378:metabolism*; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000378:metabolism; D001483:Base Sequence; D015815:Cell Adhesion Molecules / Q000096:biosynthesis* / Q000378:metabolism; D003001:Cloning, Molecular; D017124:Conserved Sequence; D017931:DNA Primers; D000939:Epitopes / Q000032:analysis; D005656:Fungal Proteins / Q000096:biosynthesis* / Q000378:metabolism; D005800:Genes, Fungal; D005838:Genotype; D008830:Microbodies / Q000378:metabolism*; D016253:Microscopy, Immunoelectron; D008969:Molecular Sequence Data; D016296:Mutagenesis; D015388:Organelles / Q000378:metabolism; D000074437:Peroxisomal Targeting Signal 2 Receptor; D010957:Plasmids; D016133:Polymerase Chain Reaction; D014176:Protein Biosynthesis; D018160:Receptors, Cytoplasmic and Nuclear; D012091:Repetitive Sequences, Nucleic Acid; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins; D014158:Transcription, Genetic
chemical_list: D001426:Bacterial Proteins; D015815:Cell Adhesion Molecules; D017931:DNA Primers; D000939:Epitopes; D005656:Fungal Proteins; C082902:PEX7 protein, S cerevisiae; D000074437:Peroxisomal Targeting Signal 2 Receptor; D018160:Receptors, Cytoplasmic and Nuclear; D029701:Saccharomyces cerevisiae Proteins; D000101:Acetyl-CoA C-Acetyltransferase