The Hansenula polymorpha PER1 gene is essential for peroxisome biogenesis and encodes a peroxisomal matrix protein with both carboxy- and amino-terminal targeting signals.
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 support.
Waterham
H R
HR
Department of Microbiology, University of Groningen, Haren, The Netherlands.
Titorenko
V I
VI
Haima
P
P
Cregg
J M
JM
Harder
W
W
Veenhuis
M
M
D000595:Amino Acid Sequence; D001483:Base Sequence; D015152:Blotting, Northern; D003001:Cloning, Molecular; D017931:DNA Primers; D004271:DNA, Fungal / Q000737:chemistry / Q000378:metabolism; D005656:Fungal Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D005800:Genes, Fungal; D005816:Genetic Complementation Test; D008830:Microbodies / Q000502:physiology* / Q000648:ultrastructure; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D016296:Mutagenesis; D016366:Open Reading Frames; D010641:Phenotype; D010843:Pichia / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure*; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism; D015183:Restriction Mapping; D015398:Signal Transduction; D001618:beta-Lactamases / Q000032:analysis / Q000378:metabolism
D017931:DNA Primers; D004271:DNA, Fungal; D005656:Fungal Proteins; D012333:RNA, Messenger; D001618:beta-Lactamases