• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of embojLink to Publisher's site
EMBO J. Dec 15, 1997; 16(24): 7326–7341.
PMCID: PMC1170333

Overexpression of Pex15p, a phosphorylated peroxisomal integral membrane protein required for peroxisome assembly in S.cerevisiae, causes proliferation of the endoplasmic reticulum membrane.

Abstract

We have cloned PEX15 which is required for peroxisome biogenesis in Saccharomyces cerevisiae. pex15Delta cells are characterized by the cytosolic accumulation of peroxisomal matrix proteins containing a PTS1 or PTS2 import signal, whereas peroxisomal membrane proteins are present in peroxisomal remnants. PEX15 encodes a phosphorylated, integral peroxisomal membrane protein (Pex15p). Using multiple in vivo methods to determine the topology, Pex15p was found to be a tail-anchored type II (Ncyt-Clumen) peroxisomal membrane protein with a single transmembrane domain near its carboxy-terminus. Overexpression of Pex15p resulted in impaired peroxisome assembly, and caused profound proliferation of the endoplasmic reticulum (ER) membrane. The lumenal carboxy-terminal tail of Pex15p protrudes into the lumen of these ER membranes, as demonstrated by its O-glycosylation. Accumulation in the ER was also observed at an endogenous expression level when Pex15p was fused to the N-terminus of mature invertase. This resulted in core N-glycosylation of the hybrid protein. The lumenal C-terminal tail of Pex15p is essential for targeting to the peroxisomal membrane. Furthermore, the peroxisomal membrane targeting signal of Pex15p overlaps with an ER targeting signal on this protein. These results indicate that Pex15p may be targeted to peroxisomes via the ER, or to both organelles.

Full Text

The Full Text of this article is available as a PDF (880K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Baerends RJ, Rasmussen SW, Hilbrands RE, van der Heide M, Faber KN, Reuvekamp PT, Kiel JA, Cregg JM, van der Klei IJ, Veenhuis M. The Hansenula polymorpha PER9 gene encodes a peroxisomal membrane protein essential for peroxisome assembly and integrity. J Biol Chem. 1996 Apr 12;271(15):8887–8894. [PubMed]
  • Bodnar AG, Rachubinski RA. Characterization of the integral membrane polypeptides of rat liver peroxisomes isolated from untreated and clofibrate-treated rats. Biochem Cell Biol. 1991 Aug;69(8):499–508. [PubMed]
  • Diestelkötter P, Just WW. In vitro insertion of the 22-kD peroxisomal membrane protein into isolated rat liver peroxisomes. J Cell Biol. 1993 Dec;123(6 Pt 2):1717–1725. [PMC free article] [PubMed]
  • Distel B, Erdmann R, Gould SJ, Blobel G, Crane DI, Cregg JM, Dodt G, Fujiki Y, Goodman JM, Just WW, et al. A unified nomenclature for peroxisome biogenesis factors. J Cell Biol. 1996 Oct;135(1):1–3. [PMC free article] [PubMed]
  • Dyer JM, McNew JA, Goodman JM. The sorting sequence of the peroxisomal integral membrane protein PMP47 is contained within a short hydrophilic loop. J Cell Biol. 1996 Apr;133(2):269–280. [PMC free article] [PubMed]
  • Elgersma Y, Tabak HF. Proteins involved in peroxisome biogenesis and functioning. Biochim Biophys Acta. 1996 Oct 29;1286(3):269–283. [PubMed]
  • Elgersma Y, van den Berg M, Tabak HF, Distel B. An efficient positive selection procedure for the isolation of peroxisomal import and peroxisome assembly mutants of Saccharomyces cerevisiae. Genetics. 1993 Nov;135(3):731–740. [PMC free article] [PubMed]
  • Elgersma Y, van Roermund CW, Wanders RJ, Tabak HF. Peroxisomal and mitochondrial carnitine acetyltransferases of Saccharomyces cerevisiae are encoded by a single gene. EMBO J. 1995 Jul 17;14(14):3472–3479. [PMC free article] [PubMed]
  • Elgersma Y, Kwast L, Klein A, Voorn-Brouwer T, van den Berg M, Metzig B, America T, Tabak HF, Distel B. The SH3 domain of the Saccharomyces cerevisiae peroxisomal membrane protein Pex13p functions as a docking site for Pex5p, a mobile receptor for the import PTS1-containing proteins. J Cell Biol. 1996 Oct;135(1):97–109. [PMC free article] [PubMed]
  • Elgersma Y, Vos A, van den Berg M, van Roermund CW, van der Sluijs P, Distel B, Tabak HF. Analysis of the carboxyl-terminal peroxisomal targeting signal 1 in a homologous context in Saccharomyces cerevisiae. J Biol Chem. 1996 Oct 18;271(42):26375–26382. [PubMed]
  • Erdmann R, Blobel G. Identification of Pex13p a peroxisomal membrane receptor for the PTS1 recognition factor. J Cell Biol. 1996 Oct;135(1):111–121. [PMC free article] [PubMed]
  • Erdmann R, Kunau WH. A genetic approach to the biogenesis of peroxisomes in the yeast Saccharomyces cerevisiae. Cell Biochem Funct. 1992 Sep;10(3):167–174. [PubMed]
  • Fujiki Y, Fowler S, Shio H, Hubbard AL, Lazarow PB. Polypeptide and phospholipid composition of the membrane of rat liver peroxisomes: comparison with endoplasmic reticulum and mitochondrial membranes. J Cell Biol. 1982 Apr;93(1):103–110. [PMC free article] [PubMed]
  • Fujiki Y, Rachubinski RA, Lazarow PB. Synthesis of a major integral membrane polypeptide of rat liver peroxisomes on free polysomes. Proc Natl Acad Sci U S A. 1984 Nov;81(22):7127–7131. [PMC free article] [PubMed]
  • Gentzsch M, Tanner W. The PMT gene family: protein O-glycosylation in Saccharomyces cerevisiae is vital. EMBO J. 1996 Nov 1;15(21):5752–5759. [PMC free article] [PubMed]
  • Glover JR, Andrews DW, Rachubinski RA. Saccharomyces cerevisiae peroxisomal thiolase is imported as a dimer. Proc Natl Acad Sci U S A. 1994 Oct 25;91(22):10541–10545. [PMC free article] [PubMed]
  • Gould SG, Keller GA, Subramani S. Identification of a peroxisomal targeting signal at the carboxy terminus of firefly luciferase. J Cell Biol. 1987 Dec;105(6 Pt 2):2923–2931. [PMC free article] [PubMed]
  • Gould SJ, Keller GA, Schneider M, Howell SH, Garrard LJ, Goodman JM, Distel B, Tabak H, Subramani S. Peroxisomal protein import is conserved between yeast, plants, insects and mammals. EMBO J. 1990 Jan;9(1):85–90. [PMC free article] [PubMed]
  • Gould SJ, Kalish JE, Morrell JC, Bjorkman J, Urquhart AJ, Crane DI. Pex13p is an SH3 protein of the peroxisome membrane and a docking factor for the predominantly cytoplasmic PTs1 receptor. J Cell Biol. 1996 Oct;135(1):85–95. [PMC free article] [PubMed]
  • Häusler T, Stierhof YD, Wirtz E, Clayton C. Import of a DHFR hybrid protein into glycosomes in vivo is not inhibited by the folate-analogue aminopterin. J Cell Biol. 1996 Feb;132(3):311–324. [PMC free article] [PubMed]
  • Herscovics A, Orlean P. Glycoprotein biosynthesis in yeast. FASEB J. 1993 Apr 1;7(6):540–550. [PubMed]
  • Hettema EH, van Roermund CW, Distel B, van den Berg M, Vilela C, Rodrigues-Pousada C, Wanders RJ, Tabak HF. The ABC transporter proteins Pat1 and Pat2 are required for import of long-chain fatty acids into peroxisomes of Saccharomyces cerevisiae. EMBO J. 1996 Aug 1;15(15):3813–3822. [PMC free article] [PubMed]
  • Heyman JA, Monosov E, Subramani S. Role of the PAS1 gene of Pichia pastoris in peroxisome biogenesis. J Cell Biol. 1994 Dec;127(5):1259–1273. [PMC free article] [PubMed]
  • Imanaka T, Shiina Y, Takano T, Hashimoto T, Osumi T. Insertion of the 70-kDa peroxisomal membrane protein into peroxisomal membranes in vivo and in vitro. J Biol Chem. 1996 Feb 16;271(7):3706–3713. [PubMed]
  • Johnson LM, Bankaitis VA, Emr SD. Distinct sequence determinants direct intracellular sorting and modification of a yeast vacuolar protease. Cell. 1987 Mar 13;48(5):875–885. [PubMed]
  • Jones AL, Fawcett DW. Hypertrophy of the agranular endoplasmic reticulum in hamster liver induced by phenobarbital (with a review on the functions of this organelle in liver). J Histochem Cytochem. 1966 Mar;14(3):215–232. [PubMed]
  • Klein P, Kanehisa M, DeLisi C. The detection and classification of membrane-spanning proteins. Biochim Biophys Acta. 1985 May 28;815(3):468–476. [PubMed]
  • Kragler F, Langeder A, Raupachova J, Binder M, Hartig A. Two independent peroxisomal targeting signals in catalase A of Saccharomyces cerevisiae. J Cell Biol. 1993 Feb;120(3):665–673. [PMC free article] [PubMed]
  • Kutay U, Ahnert-Hilger G, Hartmann E, Wiedenmann B, Rapoport TA. Transport route for synaptobrevin via a novel pathway of insertion into the endoplasmic reticulum membrane. EMBO J. 1995 Jan 16;14(2):217–223. [PMC free article] [PubMed]
  • Kyte J, Doolittle RF. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. [PubMed]
  • Lazarow PB, Fujiki Y. Biogenesis of peroxisomes. Annu Rev Cell Biol. 1985;1:489–530. [PubMed]
  • Lüers G, Hashimoto T, Fahimi HD, Völkl A. Biogenesis of peroxisomes: isolation and characterization of two distinct peroxisomal populations from normal and regenerating rat liver. J Cell Biol. 1993 Jun;121(6):1271–1280. [PMC free article] [PubMed]
  • McNew JA, Goodman JM. An oligomeric protein is imported into peroxisomes in vivo. J Cell Biol. 1994 Dec;127(5):1245–1257. [PMC free article] [PubMed]
  • Nasmyth KA, Tatchell K. The structure of transposable yeast mating type loci. Cell. 1980 Mar;19(3):753–764. [PubMed]
  • Ohno S, Fujii Y. Three-dimensional and histochemical studies of peroxisomes in cultured hepatocytes by quick-freezing and deep-etching method. Histochem J. 1990 Mar;22(3):143–154. [PubMed]
  • Parrish ML, Sengstag C, Rine JD, Wright RL. Identification of the sequences in HMG-CoA reductase required for karmellae assembly. Mol Biol Cell. 1995 Nov;6(11):1535–1547. [PMC free article] [PubMed]
  • Rehling P, Marzioch M, Niesen F, Wittke E, Veenhuis M, Kunau WH. The import receptor for the peroxisomal targeting signal 2 (PTS2) in Saccharomyces cerevisiae is encoded by the PAS7 gene. EMBO J. 1996 Jun 17;15(12):2901–2913. [PMC free article] [PubMed]
  • Rose MD, Novick P, Thomas JH, Botstein D, Fink GR. A Saccharomyces cerevisiae genomic plasmid bank based on a centromere-containing shuttle vector. Gene. 1987;60(2-3):237–243. [PubMed]
  • Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. [PMC free article] [PubMed]
  • Skorin C, Soto U, Necochea C, Leighton F. Protein phosphorylation in peroxisomes. Biochem Biophys Res Commun. 1986 Oct 15;140(1):188–194. [PubMed]
  • Subramani S. Protein translocation into peroxisomes. J Biol Chem. 1996 Dec 20;271(51):32483–32486. [PubMed]
  • Subramani S. PEX genes on the rise. Nat Genet. 1997 Apr;15(4):331–333. [PubMed]
  • Swinkels BW, Gould SJ, Bodnar AG, Rachubinski RA, Subramani S. A novel, cleavable peroxisomal targeting signal at the amino-terminus of the rat 3-ketoacyl-CoA thiolase. EMBO J. 1991 Nov;10(11):3255–3262. [PMC free article] [PubMed]
  • Van der Leij I, Van den Berg M, Boot R, Franse M, Distel B, Tabak HF. Isolation of peroxisome assembly mutants from Saccharomyces cerevisiae with different morphologies using a novel positive selection procedure. J Cell Biol. 1992 Oct;119(1):153–162. [PMC free article] [PubMed]
  • van Roermund CW, Elgersma Y, Singh N, Wanders RJ, Tabak HF. The membrane of peroxisomes in Saccharomyces cerevisiae is impermeable to NAD(H) and acetyl-CoA under in vivo conditions. EMBO J. 1995 Jul 17;14(14):3480–3486. [PMC free article] [PubMed]
  • van Roermund CW, van den Berg M, Wanders RJ. Localization of peroxisomal 3-oxoacyl-CoA thiolase in particles of varied density in rat liver: implications for peroxisome biogenesis. Biochim Biophys Acta. 1995 Dec 14;1245(3):348–358. [PubMed]
  • Vergères G, Yen TS, Aggeler J, Lausier J, Waskell L. A model system for studying membrane biogenesis. Overexpression of cytochrome b5 in yeast results in marked proliferation of the intracellular membrane. J Cell Sci. 1993 Sep;106(Pt 1):249–259. [PubMed]
  • Walton PA, Hill PE, Subramani S. Import of stably folded proteins into peroxisomes. Mol Biol Cell. 1995 Jun;6(6):675–683. [PMC free article] [PubMed]
  • Walworth NC, Goud B, Ruohola H, Novick PJ. Fractionation of yeast organelles. Methods Cell Biol. 1989;31:335–356. [PubMed]
  • Wiemer EA, Lüers GH, Faber KN, Wenzel T, Veenhuis M, Subramani S. Isolation and characterization of Pas2p, a peroxisomal membrane protein essential for peroxisome biogenesis in the methylotrophic yeast Pichia pastoris. J Biol Chem. 1996 Aug 2;271(31):18973–18980. [PubMed]
  • Wright R, Basson M, D'Ari L, Rine J. Increased amounts of HMG-CoA reductase induce "karmellae": a proliferation of stacked membrane pairs surrounding the yeast nucleus. J Cell Biol. 1988 Jul;107(1):101–114. [PMC free article] [PubMed]
  • Zhang JW, Lazarow PB. Peb1p (Pas7p) is an intraperoxisomal receptor for the NH2-terminal, type 2, peroxisomal targeting sequence of thiolase: Peb1p itself is targeted to peroxisomes by an NH2-terminal peptide. J Cell Biol. 1996 Feb;132(3):325–334. [PMC free article] [PubMed]

Articles from The EMBO Journal are provided here courtesy of The European Molecular Biology Organization

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

  • Gene
    Gene
    Gene links
  • GEO Profiles
    GEO Profiles
    Related GEO records
  • MedGen
    MedGen
    Related information in MedGen
  • Pathways + GO
    Pathways + GO
    Pathways, annotations and biological systems (BioSystems) that cite the current article.
  • Protein
    Protein
    Published protein sequences
  • PubMed
    PubMed
    PubMed citations for these articles
  • Substance
    Substance
    PubChem Substance links
  • Taxonomy
    Taxonomy
    Related taxonomy entry
  • Taxonomy Tree
    Taxonomy Tree

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...