The Hansenula polymorpha PER1 gene is essential for peroxisome biogenesis and encodes a peroxisomal matrix protein with both carboxy- and amino-terminal targeting signals

• Published in: The Journal of cell biology 1994-11, Vol.127 (3), p.737-749
• Main Authors: Waterham, H.R. (University of Groningen, Haren, The Netherlands), Titorenko, V.I, Haima, P, Cregg, J.M, Harder, W, Veenhuis, M
• Format: Article
• Language: English
• Subjects: Alcohols; Amino Acid Sequence; Amino acids; Base Sequence; beta-Lactamases - analysis; beta-Lactamases - metabolism; Blotting, Northern; Cell growth; Cloning, Molecular; COMPOSICION QUIMICA; COMPOSITION CHIMIQUE; DNA; DNA Primers; DNA, Fungal - chemistry; DNA, Fungal - metabolism; Fungal Proteins - biosynthesis; Fungal Proteins - genetics; Fungal Proteins - metabolism; GENE; GENES; Genes, Fungal; Genetic Complementation Test; Genomics; HANSENULA; Hansenula polymorpha; Microbodies - physiology; Microbodies - ultrastructure; Microscopy, Electron; Molecular Sequence Data; Mutagenesis; Open Reading Frames; ORGANITE CELLULAIRE; ORGANULOS CITOPLASMICOS; Oxidases; PEPTIDE; PEPTIDOS; Peroxisomes; Phenotype; Pichia; Pichia - genetics; Pichia - metabolism; Pichia - ultrastructure; PROTEINAS; PROTEINE; Proteins; Restriction Mapping; RNA, Messenger - analysis; RNA, Messenger - metabolism; SECUENCIA NUCLEICA; SEQUENCE NUCLEIQUE; Signal Transduction; Yeasts
• ISSN: 0021-9525; 1540-8140
• DOI: 10.1083/jcb.127.3.737

We describe the cloning of the Hansenula polymorpha PER1 gene and the characterization of the gene and its product, PER1p. The gene was cloned by functional complementation of a per1 mutant of H. polymorpha, which was impaired in the import of peroxisomal matrix proteins (Pim- phenotype). The DNA sequence of PER1 predicts that PER1p is a polypeptide of 650 amino acids with no significant sequence similarity to other known proteins. PER1 expression was low but significant in wild-type H. polymorpha growing on glucose and increased during growth on any one of a number of substrates which induce peroxisome proliferation. PER1p contains both a carboxy- (PTS1) and an amino-terminal (PTS2) peroxisomal targeting signal which both were demonstrated to be capable of directing bacterial beta-lactamase to the organelle. In wild-type H. polymorpha PER1p is a protein of low abundance which was demonstrated to be localized in the peroxisomal matrix. Our results suggest that the import of PER1p into peroxisomes is a prerequisite for the import of additional matrix proteins and we suggest a regulatory function of PER1p on peroxisomal protein import