Protoplasma

, Volume 235, Issue 1–4, pp 57–66 | Cite as

Peroxisomal targeting signals in green algae

Original Article

Abstract

Peroxisomal enzymatic proteins contain targeting signals (PTS) to enable their import into peroxisomes. These targeting signals have been identified as PTS1 and PTS2 in mammalian, yeast, and higher plant cells; however, no PTS2-like amino acid sequences have been observed in enzymes from the genome database of Cyanidiochyzon merolae (Bangiophyceae), a primitive red algae. In studies on the evolution of PTS, it is important to know when their sequences came to be the peroxisomal targeting signals for all living organisms. To this end, we identified a number of genes in the genome database of the green algae Chlamydomonas reinhardtii, which contains amino acid sequences similar to those found in plant PTS. In order to determine whether these sequences function as PTS in green algae, we expressed modified green fluorescent proteins (GFP) fused to these putative PTS peptides under the cauliflower mosaic virus 35S promoter. To confirm whether granular structures containing GFP–PTS fusion proteins accumulated in the peroxisomes of Closterium ehrenbergii, we observed these cells after the peroxisomes were stained with 3, 3′-diaminobenzidine. Our results confirm that the GFP–PTS fusion proteins indeed accumulated in the peroxisomes of these green algae. These findings suggest that the peroxisomal transport system for PTS1 and PTS2 is conserved in green algal cells and that our fusion proteins can be used to visualize peroxisomes in live cells.

Keywords

Closterium Green algae Peroxisomal targeting signal (PTS) Peroxisome Transport 

Notes

Acknowledgments

We thank Dr Akira Kato for providing the cDNA of pumpkin citrate synthase and Dr. Yasuo Niwa (University of Shizuoka, Shizuoka) for providing the 35S-GFP(S65T) plasmid. This work was supported by a Grant for Promotion of Niigata University Research Projects.

References

  1. Albertini M, Rehling P, Erdmann R, Girzalsky W, Kiel JA, Veenhuis M, Kunau WH (1997) Pex14 p, a peroxisomal membrane protein binding both receptors of the two PTS-dependent import pathways. Cell 89:83–92PubMedCrossRefGoogle Scholar
  2. Assaad FF, Signer ER (1990) Cauliflower mosaic virus P35S promoter activity in Escherichia coli. Mol Gen Genet 223:517–520PubMedCrossRefGoogle Scholar
  3. Baker A (1996) In vitro systems in the study of peroxisomal protein import. Experienti 52:1055–1062CrossRefGoogle Scholar
  4. Beevers H (1979) Microbodies in higher plants. Annu Rev Plant Physiol 30:159–193CrossRefGoogle Scholar
  5. Berger F, Linstead P, Dolan L, Haseloff J (1998) Stomata patterning on the hypocotyl of Arabidopsis thaliana is controlled by genes involved in the control of root epidermis patterning. Dev Biol 194:226–234PubMedCrossRefGoogle Scholar
  6. Blattner J, Swinkels B, Dorsam H, Prospero T, Subramani S, Clayton C (1992) Glycosome assembly in trypanosomes:variation in the acceptable degeneracy of a COOH-terminal microbody targeting signal. J Cell Biol 119:1129–1136PubMedCrossRefGoogle Scholar
  7. Blattner J, Dorsam H, Clayton CE (1995) Function of N-terminal import signals in trypanosome microbodies. FEBS Lett 360:310–314PubMedCrossRefGoogle Scholar
  8. Burke C, Yu XB, Marchitelli L, Davis EA, Ackerman S (1990) Transcription factor IIA of wheat and human function similarly with plant and animal viral promoters. Nucleic Acids Res 18:3611–3620PubMedCrossRefGoogle Scholar
  9. Cavalier-Smith T (1997) Cell and genome coevolution: facultative anaerobiosis, glycosomes and kinetoplastan RNA editing. Trends Genet 13:6–9PubMedCrossRefGoogle Scholar
  10. Chiu W-1, Niwa Y, Zeng W, Hirano T, Kobayashi H, Sheen J (1996) Engineered GFP as a vital reporter in plants. Curr Biol 6:325–330PubMedCrossRefGoogle Scholar
  11. Cooke R, Penon P (1990) In vitro transcription from cauliflower mosaic virus promoters by a cell-free extract from tobacco cells. Plant Mol Biol 14:391–405PubMedCrossRefGoogle Scholar
  12. De Duve CD, Baudhuin P (1966) Peroxisomes (microbodies and related particles). Physiol Rev 46:323–357PubMedGoogle Scholar
  13. De Bellis L, Nishimura M (1991) Development of enzymes of the glyoxylate cycle during senescence of pumpkin cotyledons. Plant Cell Physiol 32:555–561Google Scholar
  14. Erdmann R, Blobel G (1996) Identification of Pex13 p a peroxisomal membrane receptor for the PTS1 recognition factor. J Cell Biol 135:111–121PubMedCrossRefGoogle Scholar
  15. Faber KN, Haima P, Gietl C, Harder W, Ab G, Veenhuis M (1994) The methylotrophic yeast Hansenula polymorpha contains an inducible import pathway for peroxisomal matrix 26 proteins with an N-terminal targeting signal (PTS2 proteins). Proc Natl Acad Sci USA 91:12985–12989PubMedCrossRefGoogle Scholar
  16. Gietl C (1990) Glyoxysomal malate dehydrogenase from watermelon is synthesized with an amino-terminal transit peptide. Proc Natl Acad Sci USA 87:5773–5777PubMedCrossRefGoogle Scholar
  17. Gould SJ, Keller GA, Schneider M, Howell SH, Garrard LJ, Goodman JM, Distel B, Tabak H, Subramani S (1990) Peroxisomal protein import is conserved between yeast, plants, insects and mammals. EMBO J 9:85–90PubMedGoogle Scholar
  18. Hayashi M, Aoki M, Kondo M, Nishimura M (1997) Changes in targeting efficiencies of proteins to plant microbodies caoused by amino acid substitutions in carboxy-terminal tripeptide. Plant Cell Physiol 38:759–768PubMedGoogle Scholar
  19. Hayashi M, Toriyama K, Kondo M, Nishimura M (1998) 2,4-Dichlorophenoxybutyric acid-resistant mutants of Arabidopsis have defects in glyoxysomal fatty acid β-oxidation. Plant Cell 10:183–195Google Scholar
  20. Hijikata M, Ishii N, Kagamiyama H, Osumi T, Hashimoto T (1987) Structural analysis of cDNA for rat peroxisomal 3-ketoacyl-COA thiolase. J Biol Chem 262:8151–8158PubMedGoogle Scholar
  21. Ichimura T (1971) Sexual cell division and conjugationpapilla formation in sexual reproduction of Closterium strigosum. In: Nishizawa K (ed) Proc 7th Int Seaweed Symp. University of Tokyo Press, Tokyo, pp 208–214Google Scholar
  22. Jacob D, Lewin A, Meister B, Appel B (2002) Plant-specific promoter sequences carry elements that are recognised by the eubacterial transcription machinery. Transgenic Res 11:291–303PubMedCrossRefGoogle Scholar
  23. Jansen GA, Ofman R, Ferdinandusse S, Ijlst L, Muijsers AO, Skjeldal OH, Stokke O, Jakobs C, Besley GTN, Wraith JE, Wanders RJA (1997) Refsum disease is caused by mutations in the phytanoyl-CoA hydroxylase gene. Nat Genet 17:190–193Google Scholar
  24. Kato A, Hayashi M, Mori H, Nishimura M (1995) Molecular characterization of a glyoxysomal citarate synthase that is synthesized as a precursor of higher molecular mass in pumpkin. Plant Mol Biol 27:377–390PubMedCrossRefGoogle Scholar
  25. Kato A, Hayashi M, Kondo M, Nishimura M (1996a) Targeting and processing of a chemeric protein with the N-terminal presequence of the precursor to glyoxysomal citrate synthase. Plant Cell 8:1601–1611PubMedCrossRefGoogle Scholar
  26. Kato A, Hayashi M, Takeuchi Y, Nishimura M (1996b) cDNA cloning and expression of a gene for 3-ketoacyl-CoA thiolase in pumpkin cotyledons. Plant Mol Biol 31:843–852PubMedCrossRefGoogle Scholar
  27. Kato A, Takeda-YoshikawaY, Hayashi M, Kondo M, Hara-Nishimura I, Nishimura M (1998) Glyoxysomal malate dehydrogenase in pumpkin: cloning of a cDNA and functional analysis of its presequence. Plant Cell Physiol 39:186–195Google Scholar
  28. Kato A, Hayashi M, Nishimura M (1999) Oligomeric proteins containing N-terminal targeting signal are imported intoperoxisomes in transgenic Arabidopsis. Plant Cell Physiol 40:586–591PubMedGoogle Scholar
  29. Kehlenbeck P, Coyal A, Tolbert NE (1995) Factors affecting development of peroxisomes and glycolate metabolism among algae of different evolutionary lines of the Prasinophyceae. Plant Physiol 109:1363–1370PubMedGoogle Scholar
  30. Komori M, Rasmussen SW, Kiel JA, Baerends RJ, Cregg JM, Klei IJV (1997) The Hansenula polymorpha PEX14 gene encodes a novel peroxisomal membrane protein essential for peroxisome biogenesis. EMBO J 16:44–53PubMedCrossRefGoogle Scholar
  31. Mano S, Hayashi M, Nishimura M (1997) Light regulates alternative splicing of hydroxypyruvate reductase in pumpkin. Plant J 17:309–320CrossRefGoogle Scholar
  32. Mano S, Nakamori C, Hayashi M, Kato A, Kondo M, Nishimura M (2002) Distribution and characterization of peroxisomes in Arabidopsis by visualization with GFP: dynamic morphology and actin dependent movement. Plant Cell Physiol 43:331–341PubMedCrossRefGoogle Scholar
  33. McNew JA, Goodman JM (1996) The targeting and assembly of peroxisomal proteins: some old rules do not apply. Trends Biochem Sci 21:54–58PubMedGoogle Scholar
  34. Miyagishima S, Kuroiwa H, Kuroiwa T (2001) The timing and manner of disassembly of the apparatuses for chloroplast and mitochondrial division in the red alga Cyanidioschyzon merolae. Planta 212:517–528PubMedCrossRefGoogle Scholar
  35. Nishimura M, Yamaguchi J, Mori H, Akazawa T, Yokota S (1986) Immunocytochemical analysis shows that glyoxysomes are directly transformed to leaf peroxisomes during greening of pumpkin cotyledon. Plant Physiol 80:313–316CrossRefGoogle Scholar
  36. Nishimura M, Takeuchi Y, Bellis LD, Hara-Nishimura I (1993) Leaf peroxisomes are directy transformed to glyoxysomes during senescence of pumpkin cotyledons. Protoplasma 175:131–137CrossRefGoogle Scholar
  37. Niwa Y, Hirano T, Yoshimoto K, Shimizu M, Kobayashi H (1999) Non-invasive quantitative detection and applications of non-toxic, S65T-type green fluorescent protein in living plants. Plant J 18:455–463PubMedCrossRefGoogle Scholar
  38. Ogawa S, Hirota K (2000) Disintegration of chloroplasts during zygote maturation in Closterium ehrenbergii (Zygnematales, Chlorophyta). J Plant Sci 161:609–614CrossRefGoogle Scholar
  39. Olsen LJ, Harada JJ (1995) Peroxisome and their assembly in higher plants. Annu Rev Plant Physiol Plant Mol Biol 46:123–146CrossRefGoogle Scholar
  40. Osumi T, Tsukamoto T, Hata S, Yokota S, Miura S, Fujiki Y, Hijikata M, Miyazawa S, Hashimoto T (1991) Amino-terminal presequence of the precursor of peroxisomal 3- ketoacyl- CoA thiolase is a cleavable signal peptide for peroxisomal targeting. Biochem Biophys Res Commun 181:947–954PubMedCrossRefGoogle Scholar
  41. Pobjecky N, Rosenberg GH, Dinter-Gottlieb G, Käufer NF (1990) Expression of the betaglucuronidase gene under the control of the CaMV 35 S promoter in Schizosaccharomyces pombe. Mol Gen Genet 220:314–316PubMedCrossRefGoogle Scholar
  42. Preisig-Müller R, Kindl H (1993) Thiolase mRNA translated in vitro yields a peptide with a putative N-terminal presequence. Plant Mol Biol 22:59–66PubMedCrossRefGoogle Scholar
  43. Saito C, Ueda T, Abe H, Wada Y, Kuroiwa T, Hisada A, Furuya M, Nakano A (2002) A complex and mobile structure forms a distinct subregion within the continuous vacuolar membrane in young cotyledons of Arabidopsis. Plant J 29: 245–255Google Scholar
  44. Silverberg BA (1975a) 3,3′-Diaminobenzidine (DAB) ultrastructural cytochemistry of microbodies in Chlorogonium elongatum. Protoplasma 85:373–376PubMedCrossRefGoogle Scholar
  45. Silverberg BA (1975b) An ultrastructural and cytochemical characterization of microbodies in the green algae. Protoplasm 83:269–295CrossRefGoogle Scholar
  46. Silverberg BA, Sawa T (1974) Cytochemical localization of oxidase activities with diamino benizidine in the green algae Chlamydomonas dysosmos. Protoplasm 81:177–188CrossRefGoogle Scholar
  47. Stabenau H (1974) Verteilung von microbody-enzyme aus Chlamydomonas in Dichtegradienten. Planta 118:35–42Google Scholar
  48. Stabenau H, Säftel W (1982) Peroxisomal glycolate oxidase in the alga Mougeotia. Planta 154:165–167CrossRefGoogle Scholar
  49. Stabenau H, Winkler U, Säftel W (1984) Mitochondrial metabolism of glycolate in the alga Eremosphaera viridis. Z Pflanzenphysiol 29:1377–1388Google Scholar
  50. Subramani S (1993) Protein import into peroxisomes and biogenesis of the organelle. Annu Rev Cell Biol 9:445–478PubMedCrossRefGoogle Scholar
  51. Swinkels BW, Gould SJ, Bodnar AG, Rachubinski RA, Subramani S (1991) A novel, cleavable peroxisomal targeting signal at the amino-terminus of the rat 3-ketoacyl-CoA thiolase. EMBO J 10:3255–3262PubMedGoogle Scholar
  52. Titus DE, Becker WM (1985) Investigation of the glyoxysome-peroxisome transition in germinating cucumber cotyledons using double-label immunoelectron microscopy. J Cell Biol 101:1288–1299PubMedCrossRefGoogle Scholar
  53. Tourte M (1972) Mise en evidence dune activite catalasique dans les peroxysimes de Micrasterias fimbriata. Planta 105:50–59CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  1. 1.Graduate School of Science and TechnologyNiigata UniversityNinotyou, Niigata cityJapan
  2. 2.Faculty of ScienceNiigata UniversityNinotyou, Niigata cityJapan

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