Giant peroxisomes in oleic acid-induced Saccharomyces cerevisiae lacking the peroxisomal membrane protein Pmp27p.
We have purified peroxisomal membranes from Saccharomyces cerevisiae after induction of peroxisomes in oleic acid-containing media. About 30 distinct proteins could be discerned among the HPLC- and SDS-PAGE-separated proteins of the high salt-extracted peroxisomal membranes. The most abundant of these, Pmp27p, was purified and the corresponding gene PMP27 was cloned and sequenced. Its primary structure is 32% identical to PMP31 and PMP32 of the yeast Candida biodinii (Moreno, M., R. Lark, K. L. Campbell, and M. J. Goodman. 1994. Yeast. 10:1447-1457). Immunoelectron microscopic localization of Pmp27p showed labeling of the peroxisomal membrane, but also of matrix-less and matrix containing tubular membranes nearby. Electronmicroscopical data suggest that some of these tubular extensions might interconnect peroxisomes to form a peroxisomal reticulum. Cells with a disrupted PMP27 gene (delta pmp27) still grew well on glucose or ethanol, but they failed to grow on oleate although peroxisomes were still induced by transfer to oleate-containing media. The induced peroxisomes of delta pmp27 cells were fewer but considerably larger than those of wild-type cells, suggesting that Pmp27p may be involved in parceling of peroxisomes into regular quanta. delta pmp27 cells cultured in oleate-containing media form multiple buds, of which virtually all are peroxisome deficient. The growth defect of delta pmp27 cells on oleic acid appears to result from the inability to segregate the giant peroxisomes to daughter cells.
Erdmann
R
R
Laboratory of Cell Biology, Howard Hughes Medical Institute, Rockefeller University, New York 10021.
Blobel
G
G
D018528:ATP-Binding Cassette Transporters; D000101:Acetyl-CoA C-Acetyltransferase / Q000276:immunology / Q000302:isolation & purification; D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D000431:Ethanol / Q000378:metabolism; D005111:Extrachromosomal Inheritance; D005656:Fungal Proteins / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D017353:Gene Deletion; D015966:Gene Expression Regulation, Fungal; D005947:Glucose / Q000378:metabolism; D007150:Immunohistochemistry; D007425:Intracellular Membranes / Q000737:chemistry / Q000648:ultrastructure; D008565:Membrane Proteins / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D008830:Microbodies / Q000235:genetics* / Q000378:metabolism / Q000648:ultrastructure; D016253:Microscopy, Immunoelectron; D008969:Molecular Sequence Data; D019301:Oleic Acid; D009829:Oleic Acids / Q000378:metabolism* / Q000494:pharmacology; D000074401:Peroxins; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism / Q000648:ultrastructure; D029701:Saccharomyces cerevisiae Proteins; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013347:Subcellular Fractions / Q000737:chemistry / Q000648:ultrastructure
D018528:ATP-Binding Cassette Transporters; D005656:Fungal Proteins; D008565:Membrane Proteins; D009829:Oleic Acids; C092128:PEX11 protein, S cerevisiae; C070895:PEX3 protein, S cerevisiae; D000074401:Peroxins; C406793:SPF1 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D019301:Oleic Acid; D000431:Ethanol; D000101:Acetyl-CoA C-Acetyltransferase; D005947:Glucose