Titorenko, Vladimir I. and Smith, Jennifer J. and Szilard, Rachel K. and Rachubinski, Richard A.
Pex20p of the Yeast Yarrowia lipolytica Is Required for the Oligomerization of Thiolase in the Cytosol and for Its Targeting to the Peroxisome.
The Journal of Cell Biology, 142
- Published Version
Official URL: http://dx.doi.org/10.1083/jcb.142.2.403
Pex mutants are defective in peroxisome assembly. In the pex20-1 mutant strain of the yeast Yarrowia lipolytica, the peroxisomal matrix protein thiolase is mislocalized exclusively to the cytosol, whereas the import of other peroxisomal proteins is unaffected. The PEX20 gene was isolated by functional complementation of the pex20-1 strain and encodes a protein, Pex20p, of 424 amino acids (47,274 D). Despite its role in the peroxisomal import of thiolase, which is targeted by an amino-terminal peroxisomal targeting signal-2 (PTS2), Pex20p does not exhibit homology to Pex7p, which acts as the PTS2 receptor. Pex20p is mostly cytosolic, whereas 4–8% is associated with high-speed (200,000 g) pelletable peroxisomes. In the wild-type strain, all newly synthesized thiolase is associated with Pex20p in a heterotetrameric complex composed of two polypeptide chains of each protein. This association is independent of PTS2. Pex20p is required for both the oligomerization of thiolase in the cytosol and its targeting to the peroxisome. Our data suggest that monomeric Pex20p binds newly synthesized monomeric thiolase in the cytosol and promotes the formation of a heterotetrameric complex of these two proteins, which could further bind to the peroxisomal membrane. Translocation of the thiolase homodimer into the peroxisomal matrix would release Pex20p monomers back to the cytosol, thereby permitting a new cycle of binding-oligomerization-targeting-release for Pex20p and thiolase.
References:, C, P Orlean, PW Robins, CB Hirschberg(1989) Topography of glycosylation in yeast: characterization of GDP-mannose transport and lumenal guanosine diphosphatase activities in Golgi-like vesicles. Proc Natl Acad Sci USA 86:6935–6939, pmid:2476806.
Albertini, M, P Rehling, R Erdmann, W Girzalsky, JAKW Kiel, M Veenhuis, W-H Kunau(1997) Pex14p, a peroxisomal membrane protein binding both receptors of the two PTS-dependent import pathways. Cell 89:83–92, pmid:9094717.
Ausubel, F.J., R. Brent, R.E. Kingston, D.D. Moore, J.G. Seidman, J.A. Smith, and K. Struhl. 1989. In Current Protocols in Molecular Biology. Greene Publishing Associates, New York. 3.1–3.17, 13.11–13.12.
Barth, G, T Scheuber(1993) Cloning of the isocitrate lyase gene (ICL1) from Yarrowia lipolyticaand characterization of the deduced protein. Mol Gen Genet 241:422–430, pmid:8246896.
Bellion, E, JM Goodman(1987) Proton ionophores prevent assembly of a peroxisomal protein. Cell 48:165–173, pmid:3539364.
Boisramé, A, J-M Beckerich, C Gaillardin(1996) Sls1p, an endoplasmic reticulum component, is involved in the protein translocation process in the yeast Yarrowia lipolytica. J Biol Chem 271:11668–11675, pmid:8662639.
Braverman, N, G Steel, C Obie, A Moser, H Moser, SJ Gould, D Valle(1997) Human PEX7encodes the peroxisomal PTS2 receptor and is responsible for rhizomelic chondrodysplasia punctata. Nat Genet 15:369–376, pmid:9090381.
Bukau, B, T Hesterkamp, J Luirink(1996) Growing in a dangerous environment: a network of multiple targeting and folding pathways for nascent polypeptides in the cytosol. Trends Cell Biol 6:480–486, pmid:15157507.
Coligan, J.E., B.M. Dunn, H.L. Ploegh, D.W. Speicher, and P.T Wingfield. 1995. In Current Protocols in Protein Science. Wiley Interscience, New York. 10.11.1–10.11.6.
Dingwall, C, S Kandels-Lewis, B Séraphin(1995) A family of Ran binding proteins that includes nucleoporins. Proc Natl Acad Sci USA 92:7525–7529, pmid:7638224.
Distel, B, R Erdmann, SJ Gould, G Blobel, DI Crane, JM Cregg, G Dodt, Y Fujiki, JM Goodman, WW Just, JAKW Kiel, W-H Kunau, PB Lazarow, GP Mannaerts, H Moser, T Osumi, RA Rachubinski, A Roscher, S Subramani, HF Tabak, D Valle, I van der Klei, PP van Veldhoven, M Veenhuis(1996) A unified nomenclature for peroxisome biogenesis. J Cell Biol 135:1–3, pmid:8858157.
Dodt, G, SJ Gould(1996) Multiple PEXgenes are required for proper subcellular distribution and stability of Pex5p, the PTS1 receptor: evidence that PTS1 protein import is mediated by a cycling receptor. J Cell Biol 135:1763–1774, pmid:8991089.
Eitzen, GA, JD Aitchison, RK Szilard, M Veenhuis, WM Nuttley, RA Rachubinski(1995) The Yarrowia lipolytica gene PAY2encodes a 42-kDa peroxisomal integral membrane protein essential for matrix protein import and peroxisome enlargement but not for peroxisome membrane proliferation. J Biol Chem 270:1429–1436, pmid:7836411.
Eitzen, GA, VI Titorenko, JJ Smith, M Veenhuis, RK Szilard, RA Rachubinski(1996) The Yarrowia lipolytica gene PAY5encodes a peroxisomal integral membrane protein homologous to the mammalian peroxisome assembly factor PAF-1. J Biol Chem 271:20300–20306, pmid:8702763.
Eitzen, GA, RK Szilard, RA Rachubinski(1997) Enlarged peroxisomes are present in oleic acid-grown Yarrowia lipolytica overexpressing the PEX16gene encoding an intraperoxisomal peripheral membrane peroxin. J Cell Biol 137:1265–1278, pmid:9182661.
Elgersma, Y, A Vos, M van den Berg, CW van Roermund, P van der Sluijs, B Distel, HF Tabak(1996) Analysis of the carboxyl-terminal peroxisomal targeting signal 1 in a homologous context in Saccharomyces cerevisiae. J Biol Chem 271:26375–26382, pmid:8824293.
Erdmann, R, M Veenhuis, W-H Kunau(1997) Peroxisomes: organelles at the crossroads. Trends Cell Biol 7:400–407, pmid:17708989.
Elgersma, Y, M Elgersma-Hooisma, T Wenzel, JM McCaffery, MG Farquhar, S Subramani(1998) A mobile PTS2 receptor for peroxisomal protein import in Pichia pastoris. J Cell Biol 140:807–820, pmid:9472033.
Franzusoff, A, J Rothblatt, R Schekman(1991) Analysis of polypeptide transit through yeast secretory pathway. Methods Enzymol 194:662–674, pmid:2005814.
Frydman, J, F-U Hartl(1996) Principles of chaperone-assisted protein folding: differences between in vitro and in vivo mechanisms. Science 272:1497–1502, pmid:8633246.
Frydman, J, E Nimmesgern, K Ohtsuka, F-U Hartl(1994) Folding of nascent polypeptide chains in a high molecular mass assembly with molecular chaperones. Nature 370:111–117, pmid:8022479.
Glover, JR, DW Andrews, RA Rachubinski(1994) a. Saccharomyces cerevisiaeperoxisomal thiolase is imported as a dimer. Proc Natl Acad Sci USA 91:10541–10545, pmid:7937990.
Glover, JR, DW Andrews, S Subramani, RA Rachubinski(1994b) Mutagenesis of the amino targeting signal of Saccharomyces cerevisiae3-ketoacyl-CoA thiolase reveals conserved amino acids required for import into peroxisomes in vivo. J Biol Chem 269:7558–7563, pmid:8125978.
Goodman, JM, SB Tramp, H Hang, M Veenhuis(1990) Peroxisomes induced in Candida Badenby methanol, oleic acid and D-alanine vary in metabolic function but share common integral membrane proteins. J Cell Sci 97:193–204, pmid:2258389.
Hartl, F-U(1996) Molecular chaperones in cellular protein folding. Nature 381:571–580, pmid:8637592.
Hendrick, JP, F-U Hartl(1993) Molecular chaperone functions of heat-shock proteins. Annu Rev Biochem 62:349–384, pmid:8102520.
Häusler, T, Y Stierhof, E Wirtz, C Clayton(1996) Import of DHFR hybrid protein into glycosomes in vivo is not inhibited by the folate-analogue aminopterin. J Cell Biol 132:311–324, pmid:8636210.
Horwich, AL, KR Willison(1993) Protein folding in the cell: functions of two families of molecular chaperone, hsp60 and TF55-TCP1. Phil Trans R Soc Lond 339:313–326, pmid:8098536.
Kunau, W.-H. 1998. Peroxisome biogenesis: from yeast to man. Curr. Opin. Microbiol. In press.
Kyhse-Andersen, J(1984) Electroblotting of multiple gels: a simple apparatus without buffer tank for rapid transfer of proteins from polyacrylamide to nitrocellulose. J Biochem Biophys Methods 10:203–209, pmid:6530509.
Laemmli, UK(1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685, pmid:5432063.
Lanzetta, PA, LJ Alvarez, PS Reinach, OA Candia(1979) An improved assay for nanomole amounts of inorganic phosphate. Anal Biochem 100:95–97, pmid:161695.
Lee, MS, RT Mullen, RN Trelease(1997) Oilseed isocitrate lyases lacking their essential type I peroxisomal targeting signal are piggybacked to glyoxysomes. Plant Cell 9:185–197, pmid:9061950.
Leiper, JM, PB Oatey, CJ Danpure(1996) Inhibition of alanine:glyoxylate aminotransferase 1 dimerization is a prerequisite for its peroxisome-to-mitochondrion mistargeting in primary hyperoxaluria type 1. J Cell Biol 135:939–951, pmid:8922378.
Lopez, MC, J-M Nicaud, HB Skinner, C Vergnolle, JC Kader, VA Bankaitis, C Gaillardin(1994) A phosphatidylinositol/phosphatidylcholine transfer protein is required for differentiation of the dimorphic yeast Yarrowia lipolyticafrom the yeast to the mycelial form. J Cell Biol 125:113–127, pmid:8138566.
Marshall, PA, JM Dyer, ME Quick, JM Goodman(1996) Redox-sensitive homodimerization of Pex11p: a proposed mechanism to regulate peroxisomal division. J Cell Biol 135:123–137, pmid:8858168.
Marzioch, M, R Erdmann, M Veenhuis, W-H Kunau(1994) PAS7encodes a novel yeast member of the WD-40 protein family essential for import of 3-oxoacyl-CoA thiolase, a PTS2-containing protein, into peroxisomes. EMBO (Eur Mol Biol Organ) J 13:4908–4918, pmid:7957058.
McNew, JA, JM Goodman(1994) An oligomeric protein is imported into peroxisomes in vivo. J Cell Biol 127:1245–1257, pmid:7962087.
McNew, JA, JM Goodman(1996) The targeting and assembly of peroxisomal proteins: some old rules do not apply. Trends Biochem Sci 21:54–58, pmid:8851661.
Motley, A, E Hettema, EM Hogenhout, P Brites, ALMA ten Asbroek, FA Wijburg, F Baas, H Heijmans, HF Tabak, RJA Wanders, B Distel(1997) Rhizomelic chondrodysplasia punctata is a peroxisomal targeting disease caused by a non-functional PTS2 receptor. Nat Genet 15:377–380, pmid:9090382.
Nicaud, J.-M., A. Le Clainche, M.-T. Le Dall, H. Wang, and C. Gaillardin. 1998. Yarrowia lipolytica, a yeast model for the genetic studies of hydroxy fatty acids biotransformation into lactones. J. Mol. Catal. B Enzymatic. 151: In press.
Nuttley, WM, AM Brade, C Gaillardin, GA Eitzen, JR Glover, JD Aitchison, RA Rachubinski(1993) Rapid identification and characterization of peroxisomal assembly mutants in Yarrowia lipolytica. Yeast 9:507–517.
Pause, B, P Diestelkötter, H Heid, WW Just(1997) Cytosolic factors mediate protein insertion into the peroxisomal membrane. FEBS (Fed Eur Biochem Soc) Lett 414:95–98, pmid:9305739.
Purdue, PE, PB Lazarow(1994) Peroxisomal biogenesis: multiple pathways of protein import. J Biol Chem 269:30065–30068, pmid:7982905.
Purdue, PE, JW Zhang, M Skoneczny, PB Lazarow(1997) Rhizomelic chondrodysplasia punctata is caused by deficiency of human Pex7p, a homologue of the yeast PTS2 receptor. Nat Genet 15:381–384, pmid:9090383.
Rachubinski, RA, S Subramani(1995) How proteins penetrate peroxisomes. Cell 83:525–528, pmid:7585954.
Radu, A, MS Moore, G Blobel(1995) The peptide repeat domain of nucleoporin Nup98 as a docking site in transport across the nuclear pore complex. Cell 81:215–222, pmid:7736573.
Rehling, P, M Marzioch, F Niesen, E Wittke, M Veenhuis, W-H Kunau(1996) The import receptor for the peroxisomal targeting signal 2 (PTS2) in Saccharomyces cerevisiae is encoded by the PAS7gene. EMBO (Eur Mol Biol Organ) J 15:2901–2913, pmid:8670791.
Roberts, CJ, CK Raymond, CT Yamashiro, TH Stevens(1991) Methods for studying the yeast vacuole. Methods Enzymol 194:644–661, pmid:1706462.
Sanders, SL, KM Whitfield, JP Vogel, MD Rose, RW Schekman(1992) Sec61p and BiP directly facilitate polypeptide translocation into the ER. Cell 69:353–365, pmid:1568250.
Schatz, G, B Dobberstein(1996) Common principles of protein translocation across membranes. Science 271:1519–1526, pmid:8599107.
Smith, JJ, RK Szilard, M Marelli, RA Rachubinski(1997) The peroxin Pex17p of the yeast Yarrowia lipolyticais associated peripherally with the peroxisomal membrane and is required for the import of a subset of matrix proteins. Mol Cell Biol 17:2511–2520, pmid:9111320.
Subramani, S(1998) Components involved in peroxisome import, biogenesis, proliferation, turnover, and movement. Pharmacol Rev 78:171–188.
Szilard, RK, VI Titorenko, M Veenhuis, RA Rachubinski(1995) Pay32p of the yeast Yarrowia lipolyticais an intraperoxisomal component of the matrix protein translocation machinery. J Cell Biol 131:1453–1469, pmid:8522603.
Thieringer, R, H Shio, YS Han, G Cohen, PB Lazarow(1991) Peroxisomes in Saccharomyces cerevisiae: immunofluorescence analysis and import of catalase A into isolated peroxisomes. Mol Cell Biol 11:510–522, pmid:1986244.
Titorenko, VI, RA Rachubinski(1998) Mutants of the yeast Yarrowia lipolyticadefective in protein exit from the endoplasmic reticulum are also defective in peroxisome biogenesis. Mol Cell Biol 18:2789–2803, pmid:9566898.
Titorenko, VI, GA Eitzen, RA Rachubinski(1996) Mutations in the PAY5 gene of the yeast Yarrowia lipolyticacause the accumulation of multiple subpopulations of peroxisomes. J Biol Chem 271:20307–20314, pmid:8702764.
Titorenko, VI, DM Ogrydziak, RA Rachubinski(1997) Four distinct secretory pathways serve protein secretion, cell surface growth, and peroxisome biogenesis in the yeast Yarrowia lipolytica. Mol Cell Biol 17:5210–5226, pmid:9271399.
Walton, PA, M Wendland, S Subramani, RA Rachubinski, WJ Welch(1994) Involvement of 70-kD heat-shock proteins in peroxisomal import. J Cell Biol 125:1037–1046, pmid:8195287.
Walton, PA, PE Hill, S Subramani(1995) Import of stably folded proteins into peroxisomes. Mol Biol Cell 6:675–683, pmid:7579687.
Waterham, HR, KA Russell, Y de Vries, JM Cregg(1997) Peroxisomal targeting, import, and assembly of alcohol oxidase in Pichia pastoris. J Cell Biol 139:1419–1431, pmid:9396748.
Zhang, JW, C Luckey, PB Lazarow(1993) Three peroxisome protein packaging pathways suggested by selective permeabilization of yeast mutants defective in peroxisome biogenesis. Mol Biol Cell 4:1351–1359, pmid:7909460.
Zhang, JW, PB Lazarow(1995) PEB1 (PAS7) in Saccharomyces cerevisiaeencodes a hydrophilic, intra-peroxisomal protein that is a member of the WD repeat family and is essential for the import of thiolase into peroxisomes. J Cell Biol 129:65–80, pmid:7535304.
Zhang, JW, PB Lazarow(1996) Peb1p (Pas7p) is an intraperoxisomal receptor for the NH2-terminal, type 2, peroxisomal targeting signal of thiolase: Peb1p itself is targeted to peroxisome by an NH2-terminal peptide. J Cell Biol 132:325–334, pmid:8636211.
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