Plant Molecular Biology

, Volume 62, Issue 3, pp 339–349 | Cite as

The Arabidopsis PPDK gene is transcribed from two promoters to produce differentially expressed transcripts responsible for cytosolic and plastidic proteins

Article

Abstract

Pyruvate orthophosphate dikinase (PPDK) is a critical enzyme for C4 photosynthesis, providing the primary acceptor for fixation of bicarbonate in mesophyll cells. Although first isolated in C4 plants, it is also present in C3 species. We report that the single gene encoding PPDK in Arabidopsis thaliana possesses two promoters, giving rise to two types of transcript. The longer transcript is generated from a promoter upstream of the first exon, while the shorter transcript is derived from a promoter found within the first intron of the longer form. Apart from 5′ untranslated regions, the presence of the first exon, and three missing codons at the start of the second exon in the longer form, the transcripts are identical. Fusions between the two forms of transcript and gfp showed that the longer transcript encodes a protein targeted to the chloroplast, that its first exon acts as a transit peptide, and that the smaller protein is cytosolic. Abundance of the shorter transcript, responsible for producing the cytosolic protein increases rapidly and specifically during extended dark and dark-induced senescence. Transcripts for both chloroplastic and cytosolic proteins were detectable in cotyledons, while in cauline leaves the transcript encoding the chloroplastic protein was most abundant. We propose that in cotyledons PPDK may be important in supplying PEP to gluconeogenesis, and in ageing leaves it allows remobilisation of nitrogen to supply reproductive tissue.

Keywords

Pyruvate orthophosphate dikinase C4 photosynthesis Arabidopsis Nitrogen remobilisation Evolution 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. An Y-Q, McDowell JM, Huang S, McKinney EC, Chambliss S, Meagher RB (1996) Strong, constitutive expression of the Arabidopsis ACT2/ACT8 actin subclass in vegetative tissues. Plant J 10:107–121PubMedCrossRefGoogle Scholar
  2. Aoyagi K, Bassham JA (1983) Pyruvate orthophosphate dikinase in wheat leaves. Plant Physiol 73:853–854PubMedGoogle Scholar
  3. Aoyagi K, Bassham JA (1984) Pyruvate orthophosphate dikinase of C3 seeds and leaves as compared to the enzyme from maize. Plant Physiol 75:387–392PubMedGoogle Scholar
  4. Aoyagi K, Bassham JA (1985) Synthesis and uptake of cytoplasmically synthesised pyruvate orthophosphate dikinase by chloroplasts. Plant Physiol 78:807–811PubMedGoogle Scholar
  5. Brown NJ, Parsley K, Hibberd JM (2005) The future of C4 research–maize, Flaveria or Cleome? Trends Plant Sci 10:215–221PubMedCrossRefGoogle Scholar
  6. Bruderer T, Wehrli C, Kohler P (1996) Cloning and characterisation of the gene encoding pyruvate phosphate dikinase from Giardia duodenalis. Mol Biochem Parasitol 77:225–233PubMedCrossRefGoogle Scholar
  7. Carroll LJ, Dunaway-Mariano D, Smith CM, Chollet R (1990) Determination of the catalytic pathway of C4-leaf pyruvate orthophosphate dikinase from maize. FEBS Lett 274:178–180PubMedCrossRefGoogle Scholar
  8. Chastain CJ, Chollet R (2003) Regulation of pyruvate orthophosphate dikinase by ADP-/Pi-dependent reversible phosphorylation in C3 and C4 plants. Plant Physiol Biochem 41:523–532CrossRefGoogle Scholar
  9. Cooper RA, Kornberg HL (1967) The direct synthesis of phosphoenolpyruvate from pyruvate by Escherichia coli. Proc R Soc Lond B Biol Sci 168:263–280PubMedCrossRefGoogle Scholar
  10. Emanuelsson O, Nielson H, von Heijne G (1999) ChloroP, a neural network-based method for predicting chloroplast transit peptides and their cleavage sites. Protein Sci 8:978–984PubMedGoogle Scholar
  11. Emanuelsson O, Nielson H, Brunak S, von Heijne G (2000) Predicting subcellular localisation of proteins based on their N-terminal amino acid sequence. J Mol Biol 300:1005–1016PubMedCrossRefGoogle Scholar
  12. Glackin CA, Grula JW (1990) Organ specific transcripts of different size and abundance derive from the same pyruvate orthophosphate dikinase gene in maize. Proc Natl Acad Sci USA 87:3004–3008PubMedCrossRefGoogle Scholar
  13. Hatch MD (1987) C4 photosynthesis: a unique blend of modified biochemistry, anatomy and ultrastructure. Biochim Biophys Acta 895:81–106Google Scholar
  14. Hatch MD, Slack CR (1968) A new enzyme for the interconversion of pyruvate and phosphopyruvate and its role in the C4 dicarboxylic acid pathway of photosynthesis. Biochem J 106:141–146PubMedGoogle Scholar
  15. Herzberg O, Chen CC, Kapadia G, McGuire M, Carroll LJ, Noh SJ, Dunaway-Matiano D (1996) Swiveling-domain mechanism for enzymatic phosphotransfer between remote reaction sites. Proc Natl Acad Sci USA 93:2652–2657PubMedCrossRefGoogle Scholar
  16. Hibberd JM, Linley PJ, Khan MS, Gray JC (1998) Transient expression of green fluorescent protein in various plastid types following micro-projectile bombardment. Plant J 16:627–632CrossRefGoogle Scholar
  17. Hibberd JM, Quick WP (2002) Characteristics of C4 photosynthesis in stems and petioles of C3 flowering plants. Nature 415:451–454PubMedCrossRefGoogle Scholar
  18. Imaizumi N, Ku MS, Ishihara K, Samejima M, Kaneko S, Matsuoka M (1997) Characterisation of the gene for pyruvate orthophosphate dikinase from rice, a C3 plant, and a comparison of structure and expression between C3 and C4 genes for this protein. Plant Mol Biol 34:701–716PubMedCrossRefGoogle Scholar
  19. Kanai R, Edwards GE (1999) The biochemistry of C4 photosynthesis. In: Sage RF Monson RK (ed) C4 plant biology. Academic Press, San Diego, pp 49–87Google Scholar
  20. Kang HG, Park S, Matsuoka M, An G (2005) White-core endosperm floury endosperm-4 in rice is generated by knockout mutations in the C4-type pyruvate orthophosphate dikinase gene (OsPPDKB). Plant J. 42:901–11PubMedCrossRefGoogle Scholar
  21. Leegood RC, Ap Rees T (1978) Phosphoenolpyruvate carboxykinase and gluconeogenesis in cotyledons of Cucurbito pepo. Biochimica et Biophysica Acta 524:208–218Google Scholar
  22. Lin J-F, Wu S-H (2004) Molecular events in senescing Arabidopsis leaves. Plant J 39:612–628PubMedCrossRefGoogle Scholar
  23. Maruyama K, Sugano S (1994) Oligo-capping: a simple method to replace the cap structure of eukaryotic mRNAs with oligoribonucleotides. Gene 138:171–174PubMedCrossRefGoogle Scholar
  24. Matsuoka M, Ozeki Y, Yamamoto N, Hirano H, Kano-Murakami Y, Tanaka Y (1988) Primary structure of maize pyruvate orthophosphate dikinase as deduced from cDNA sequence. J Biol Chem 263:11080–11083PubMedGoogle Scholar
  25. Matsuoka M, Numazawa T (1991) CIS-acting elements in the pyruvate orthophosphate dikinase gene from maize. Mol Gen Genet. 228:143–152PubMedCrossRefGoogle Scholar
  26. Moons A, Valcke R, Van Montagu M (1998) Low-oxygen stress and water deficit induce cytosolic pyruvate orthophosphate dikinase (PPDK) expression in roots of rice, a C3 plant. Plant J 15:89–98PubMedCrossRefGoogle Scholar
  27. Penfield S, Rylott EL, Gilday AD, Graham S, Larson TR, Graham IA (2004) Reserve mobilisation in the Arabidopsis endosperm fuels hypocotyl elongation in the dark, is independent of abscisic acid, and requires the PHOSPHOENOLPYRUVATE CARBOXYKINASE1 gene. Plant Cell 16:2705–2718PubMedCrossRefGoogle Scholar
  28. Pocalyko DJ, Carroll LJ, Martin BM, Babbit PC, Dunaway-Mariano D (1990) Analysis of sequence homologies in plant and bacterial pyruvate phosphate dikinase, enzyme I of the bacterial phosphoenolpyruvate: sugar phosphotransferase system and other PEP-utilising enzymes. Identification of potential catalytic and regulatory motifs. Biochemistry 29:10757–10765PubMedCrossRefGoogle Scholar
  29. Rosche E, Westhoff P (1995) Genomic structure and expression of the pyruvate, orthophosphate dikinase gene of the dicotyledonous C4 plant Flaveria trinervia (Asteraceae). Plant Mol Biol 29:663–678PubMedCrossRefGoogle Scholar
  30. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, New YorkGoogle Scholar
  31. Sheen J (1991) Molecular mechanisms underlying the differential expression of maize pyruvate, orthophosphate dikinase genes. Plant Cell 3:225–245PubMedCrossRefGoogle Scholar
  32. Simpson RH, Lambers H, Dalling MJ (1983) Nitrogen redistribution during grain growth in wheat (Triticum aestivum L.). Plant Physiol 71:7–14PubMedGoogle Scholar
  33. Small I, Peeters N, Legeai F, Lurin C (2004) Predotar: A tool for rapidly screening proteomes for N-terminal targeting sequences. Proteomics 4:1581–1590PubMedCrossRefGoogle Scholar
  34. Stewart CR, Beevers H (1967) Gluconeogenesis from amino acids in germinating castor bean endosperm and its role in transport to the embryo. Plant Physiol 41:1587–1595CrossRefGoogle Scholar
  35. Streatfield SJ, Weber A, Kinsman EA, Häusler RE, Li J, Post-Beittenmiller D, Kaiser WM, Pyke KA, Flügge UI, Chory J (1999) The phosphoenolpyruvate/ phosphate translocator is required for phenolic metabolism, palisade cell development and plastid-dependent nuclear gene expression. Plant Cell 11:1609–1621PubMedCrossRefGoogle Scholar
  36. Voll L, Häusler RE, Hecker R, Weber A, Weissenböck G, Fiene G, Waffenschmidt S, Flügge UI (2003) The phenotype of the Arabidopsis cue1 mutant is not simply caused by a general restriction of the shikimate pathway. Plant J 36:301–317PubMedCrossRefGoogle Scholar
  37. Wood HG, O’Brien WE, Michaels G (1977) Properties of carboxytransphosphorylase; pyruvate, phosphate dikinase; pyrophosphate-phosphofructokinase and pyrophosphate-acetate kinase and their roles in the metabolism of inorganic pyrophosphate. Adv Enzymol 45:85–155PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2006

Authors and Affiliations

  1. 1.Department of Plant SciencesUniversity of CambridgeCambridgeUK

Personalised recommendations