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

J Cell Biol. 1991 Sep;114(6):1167-78. doi: 10.1083/jcb.114.6.1167.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ATP-Binding Cassette Transporters*
  • Amino Acid Sequence
  • Base Sequence
  • Cloning, Molecular
  • Escherichia coli / genetics
  • Fungal Proteins / genetics*
  • Fungal Proteins / isolation & purification
  • Genes, Fungal*
  • Genetic Complementation Test
  • Genotype
  • Immunoblotting
  • Membrane Proteins / genetics*
  • Membrane Proteins / isolation & purification
  • Microbodies / physiology*
  • Microbodies / ultrastructure
  • Microscopy, Electron
  • Models, Structural
  • Molecular Sequence Data
  • Molecular Weight
  • Peroxins
  • Phenotype
  • Protein Conformation
  • Recombinant Proteins / isolation & purification
  • Restriction Mapping
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae / physiology
  • Saccharomyces cerevisiae / ultrastructure
  • Saccharomyces cerevisiae Proteins*

Substances

  • ATP-Binding Cassette Transporters
  • Fungal Proteins
  • Membrane Proteins
  • PEX3 protein, S cerevisiae
  • Peroxins
  • Recombinant Proteins
  • SPF1 protein, S cerevisiae
  • Saccharomyces cerevisiae Proteins

Associated data

  • GENBANK/M60691
  • GENBANK/M60692
  • GENBANK/M60693
  • GENBANK/M60694
  • GENBANK/M60695
  • GENBANK/M60696
  • GENBANK/M60697
  • GENBANK/S55275
  • GENBANK/X56699
  • GENBANK/X57971
  • GENBANK/X58407