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PMID: 8227124 Published · ppublish English Comparative Study Journal Article Research Support, U.S. Gov't, P.H.S.

Cloning and characterization of PAS5: a gene required for peroxisome biogenesis in the methylotrophic yeast Pichia pastoris.

The Journal of cell biology ·Vol. 123 ·No. 3 ·1993-11-00 ·Pages 535-48

Spong AP, Subramani S

Abstract

The biogenesis and maintenance of cellular organelles is of fundamental importance in all eukaryotic cells. One such organelle is the peroxisome. The establishment of a genetic system to study peroxisome biogenesis in the methylotrophic yeast Pichia pastoris has yielded many different complementation groups of peroxisomal assembly (pas) or peroxisome-deficient (per) mutants. Each appears to be deficient in functional peroxisomes. One of these mutants, pas5, has been characterized, complemented, and the gene sequenced. Ultrastructural studies show that normal peroxisomes are not present in pas5, but aberrant peroxisomal structures resembling "membranous ghosts" are frequently observed. The "peroxisome ghosts" appear to be induced and segregated to daughter cells normally. Biochemical fractionation analysis of organelles of the pas5 mutant reveals that peroxisomal matrix enzymes are induced normally but are found mostly in the cytosol. However, purification of peroxisome ghosts from the mutant shows that small amounts (< 5%) of matrix enzymes are imported. The PAS5 gene was cloned and found to encode a 127-kD protein, which contains a 200-amino acid-long region of homology with PAS1, NEM-sensitive factor (NSF), and other related ATPases. Weak homology to a yeast myosin was also observed. The gene is not essential for growth on glucose but is essential for growth on oleic acid and methanol. The role of PAS5 in peroxisome biogenesis is discussed.

MeSH Terms
Amino Acid Sequence Animals Base Sequence Cloning, Molecular Cricetinae Fungal Proteins/biosynthesis,genetics Genes, Fungal Humans Microbodies/metabolism,ultrastructure Microscopy, Electron Molecular Sequence Data Mutagenesis Pichia/genetics,metabolism Saccharomyces cerevisiae/genetics Sequence Homology, Amino Acid Swine Zellweger Syndrome/genetics,pathology
Chemicals
Fungal Proteins PAS5 protein, Pichia pastoris
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Spong A P
Department of Biology, University of California, San Diego, La Jolla 92093-0322.
Subramani S
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.123.3.535
Published
1993-11-00
Pages
535-48
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2200126
Subset
IM
Grants
NIDDK NIH HHS · DK41737 · United States
Databases
GENBANK
Z22556
External Links
PubMed source DOI: 10.1083/jcb.123.3.535 PMC Full Text View journal details
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