Abstract
Pyruvate orthophosphate dikinase (PPDK) is a key enzyme of C4 photosynthesis providing the acceptor molecule for the primary CO2 fixation in the mesophyll cells. Here we present the isolation and characterisation of the corresponding gene (termed pdk) from the C4 plant Flaveria trinervia (Asteraceae). Southern analysis indicates that in contrast to maize pdk sequences in F. trinervia are present as single copy. Sequence analysis of the entire gene reveals that its coding sequence is identical to the previous isolated PPDK-cDNA from this species. The gene spans about 13 kb and consists of 21 exons, it thus contains two additional exons compared to the maize gene. As in maize, a long intervening sequence of 6.1 kb is positioned at the boundary of the transit peptide segment and the mature protein region. Pdk transcripts accumulate abundantly in leaves, but are also detectable in stems and roots. While the leaf and stem transcripts are 3.4 kb in size and encode the chloroplastic PPDK isoform, a 3.0 kb transcript lacking the region encoding the plastidic transit peptide accumulates in roots. Thus two different transcripts can be produced from a single pdk gene most likely by use of alternative promoters and not by alternative splicing. The accumulation of the 3.4 kb transcript is under light control. Darkening leads to a drastic depletion of this transcript in both leaves and stems. Instead, the 3.0 kb transit peptide-lacking pdk transcript accumulates, but only in stems and roots, not in leaves.
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Aoyagi K, Bassham JA: Pyruvate orthophosphate dikinase in wheat leaves. Plant Physiol 73: 853–854 (1983).
Aoyagi K, Bassham JA: Pyruvate orthophosphate dikinase mRNA organ specificity in wheat and maize. Plant Physiol 76: 278–280 (1984).
Aoyagi K, Kuhlemeier C, Chua NH: Light regulation of gene expression in higher plants. Annu Rev Plant Physiol 36: 569–593 (1985).
Aoyagi K, Nakamoto H: Pyruvate,Pi dikinase in bundle sheath strands as well as in mesophyll cells in maize leaves. Plant Physiol 78: 661–664 (1985).
Aviv H, Leder P: Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci USA 69: 1408–1412 (1972).
Bruchhaus I, Tannich E: Primary structure of the pyruvate phosphate dikinase in Entamoeba histolytica. Mol Biochem Parasitol 62: 153–156 (1993).
Chen EY, Seeburg PH: Supercoil sequencing: a fast and simple method for sequencing plasmid DNA. DNA 4: 165–170 (1985).
Church GM, Gilbert W: Genomic sequencing. Proc Natl Acad Sci USA 81: 1991–1995 (1984).
Cockburn W: Stomatal mechanism as the basis of the evolution of CAM and C4 photosynthesis. Plant Cell Environ 6: 275–279 (1983).
Cole KP, Blakeley SD, Dennis DT: Structure of the gene encoding potato cytosolic pyruvate kinase. Gene 122: 255–261 (1992).
Cushman JC, Meyer G, Michalowski CB, Schmitt JM, Bohnert HJ: Salt stress leads to differential expression of two isogenes of phosphoenolpyruvate carboxylase during crassulancean acid metabolism induction in the common ice plant. Plant Cell 1: 715–725 (1989).
Darnell JE, Doolittle WF: Speculations on the early course of evolution. Proc Natl Acad Sci USA 83: 1271–1275 (1986).
Dorit RL, Gilbert W: The limited universe of exons. Curr Opin Genet Devel 1: 464–469 (1991).
Edwards GE, Ku MSB. Biochemistry of C3-C4 intermediates. In: Hatch MD, Boardman NK (eds) The Biochemistry of Plants, vol. 10, pp 275–325. Academic Press, Inc., New York (1987).
Edwards GE, Nakamoto H, Burnell JN, Hatch MD: Pyruvate,Pi dikinase and NADP-malate dehydrogenase in C4 photosynthesis: properties and mechanism of light/dark regulation. Annu Rev Plant Physiol 36: 255–286 (1985).
Edwards GE, Walker DA. C3, C4: Mechanism, and Cellular and Environmental Regulation, of Photosynthesis. Blackwell Scientific Publications, Oxford London (1983).
Ehleringer JR, Sage RF, Flanagan LB, Pearcy RW. Climate change and the evolution of C4 photosynthesis. Trends Ecol Evol 6: 95–99 (1991).
Feinberg AP, Vogelstein B: A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132: 6–13 (1983).
Glackin CA, Grula JW: Organ-specific transcripts of different size and abundance derive from the same pyruvate, orthophosphate dikinase gene in maize. Proc Natl Acad Sci USA 87: 3004–3008 (1990).
Hanley BA, Schuler MA: Plant intron sequences: evidence for distinct groups of introns. Nucl Acids Res 16: 7159–7174 (1988).
Harvey PH, Purvis A: Comparative methods for explaining adaptations. Nature 351: 619–624 (1991).
Hata S, Matsuoka M: Immunological studies on pyruvate orthophosphate dikinase in C3 plants. Plant Cell Physiol 28: 635–641 (1987).
Hatch MD: C4 photosynthesis: a unique blend of modified biochemistry, anatomy and ultrastructure. Biochim Biophys Acta 895: 81–106 (1987).
Hattersley PW. Variations in photosynthetic pathway. In: Soderstrom TR, Hilu KW, Campbell CS, Barkworth ME (eds) Grass Systematics and Evolution, pp. 49–64. Smithonian Institution Press, Washington (1987).
Helentjaris T, Weber D, Wright S: Identification of the genomic locations of duplicate nucleotide sequences in maize by analysis of restriction fragment length polymorphisms. Genetics 118: 353–363 (1988).
Henikoff S: Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene 28: 351–359 (1984).
Hermans J, Westhoff P: Analysis of expression and evolutionary relationships of phosphoenolpyruvate carboxylase genes in Flaveria trinervia (C4) and F. pringlei (C3). Mol Gen Genet 224: 459–468 (1990).
Hermans J, Westhoff P: Homologous genes for the C4 isoform of phospheenolpyruvate carboxylase in a C3-and a C4-Flaveria species. Mol Gen Genet 234: 275–284 (1992).
Hudspeth RL, Glackin CA, Bonner J, Grula JW: Genomic and cDNA clones for maize phosphoenolpyruvate carboxylase and pyruvate,orthophosphate dikinase: expression of different gene-family members in leaves and roots. Proc Natl Acad Sci USA 83: 2884–2888 (1986).
Höfer MU, Santore UJ, Westhoff P: Differential accumulation of the 10-, 16- and 23-kDa peripheral components of the water-splitting complex of photosystem II in mesophyll and bundle-sheath chloroplasts of the dicotyledonous C4 plant Flaveria trinervia (Spreng.) C. Mohr. Planta 186: 304–312 (1992).
Lepiniec L, Keryer E, Philippe H, Gadal P, Crétin C: Sorghum phosphoenolpyruvate carboxylase gene family: Structure, function and molecular evolution. Plant Mol Biol 21: 487–502 (1993).
Lipka B, Steinmüller K, Rosche E, Börsch D, Westhoff P. The C3 plant Flaveria pringlei contains a plastidic NADP-malic enzyme which is orthologous to the C4 iso-form of the C4 plant F. trinervia. Plant Mol Biol 26: 1775–1783 (1994).
Lou H, McCullough AJ, Schuler MA: 3′ Splice site selection in dicot plant nuclei is position dependent. Mol Cell Biol 13: 4485–4493 (1993).
Matsuoka M: Structure, genetic mapping, and expression of the gene for pyruvate, orthophosphate dikinase from maize. J Biol Chem 265: 16772–16777 (1990).
Matsuoka M, Ozeki Y, Yamamoto N, Hirano H, Kano-Murakami Y, Tanaka Y: Primary structure of maize pyruvate, orthophosphate dikinase as deduced from cDNA sequence. J Biol Chem 263: 11080–11083 (1988).
Moore PD: Evolution of photosynthetic pathways in flowering plants. Nature 295: 647–648 (1982).
Norris SR, Meyer SE, Callis J: The intron of Arabidopsis thaliana polyubiquitin genes is conserved in location and is a quantitative determinant of chimeric gene expression. Plant Mol Biol 21: 895–906 (1993).
Ohta T: Multigene families and the evolution of complexity. J Mol Evol 33: 34–41 (1991).
Oswald A, Streubel M, Ljungberg U, Hermans J, Eskins K, Westhoff P: Differential biogenesis of photosystem II in mesophyll and bundle sheath cells of NADP malic enzyme-type C4 plants. A comparative protein and RNA analysis. Eur J Biochem 190: 185–194 (1990).
Palmer JD, Logsdon JM: The recent origin of introns. Curr Opin Genet Devel 1: 470–477 (1991).
Pokalyco DJ, Carroll LJ, Martin MB, Babbitt PC, Cunaway-Mariano D: Analysis of sequence homologies in plant and bacterial Pyruvate Phosphate Dikinase, enzyme I of the bacterial phosphoenolpyruvate: sugar phosphotransferase system and other PEP-utilizing enzymes. Identification of potential catalytic and regulatory motifs. Biochemistry 29: 10757–10765 (1990).
Powell AM: Systematics of Flaveria (Flaveriinae-Asteraceae). Ann Mo Bot Gard 65: 590–636 (1978).
Raghavendra AS: Characteristics of plant species intermediate between C3 and C4 pathways of photosynthesis: their focus of mechanism and evolution of C4 syndrome. Photosynthetica 14: 271–283 (1980).
Rosche E, Streubel M, Westhoff P: Primary structure of the photosynthetic pyruvate orthophosphate dikinase of the C3 plant Flaveria pringlei and expression analysis of pyruvate orthophosphate dikinase sequences in C3, C3-C4 and C4 Flaveria species. Plant Mol Biol 26: 763–769 (1994).
Rosche E, Westhoff P: Primary structure of pyruvate, orthophosphate dikinase in the dicotyledonous C4 plant Flaveria trinervia. FEBS Lett 273: 116–121 (1990).
Sambrook J, Fritsch EF, Maniatis T: Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1989).
Schäffner AR, Sheen J: Maize C4 photosynthesis involves differential regulation of phosphoenolpyruvate carboxylase genes. Plant J 2: 221–232 (1992).
Sheen J: Molecular mechanisms underlying the differential expression of maize pyruvate, orthophosphate dikinase genes. Plant Cell 3: 225–245 (1991).
Steinmüller K, Batschauer A, Apel K: Tissue-specific and light-dependent changes of chromatin organization in barley (Hordeum vulgare). Eur J Biochem 158: 519–525 (1986).
Stockhaus J, Poetsch W, Steinmüller K, Westhoff P: Evolution of the C4 phosphoenolpyruvate carboxylase promoter of the C4 dicot Flaveria trinervia: an expression analysis in the C3 plant tobacco. Mol Gen Genet 245: 286–293 (1994).
Tabor S, Richardson CC: Effect of manganese ions on the incorporation of dideoxynucleotides by bacteriophage T7 DNA polymerase and Escherichia coli DNA polymerase. I. Proc Natl Acad Sci USA 86: 4076–4080 (1989).
Thompson WF, White MJ: Physiological and molecular studies of light-regulated nuclear genes in higher plants. Annu Rev Plant Physiol Plant Mol Biol 42: 423–466 (1991).
Tobin EM, Silverthorne J: Light regulation of gene expression in higher plants. Annu Rev Plant Physiol 36: 569–593 (1985).
Westhoff P, Herrmann RG: Complex RNA maturation in chloroplasts. The psbB operon from spinach. Eur J Biochem 171: 551–564 (1988).
Westhoff P, Offermann-Steinhard K, Höfer M, Eskins K, Oswald A, Streubel M: Differential accumulation of plastid transcripts encoding photosystem II components in the mesophyll and bundle-sheath cells of monocotyledonous NADP-malic enzyme-type C4 plants. Planta 184: 377–388 (1991).
Whitkus R, Doebley J, Lee M: Comparative genome mapping of sorghum and maize. Genetics 132: 1119–1130 (1992).
Wilson AC, Carlson SS, White TJ: Biochemical evolution. Annu Rev Biochem 46: 573–639 (1977).
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Rosche, E., Westhoff, P. Genomic structure and expression of the pyruvate, orthophosphate dikinase gene of the dicotyledonous C4 plant Flaveria trinervia (Asteraceae). Plant Mol Biol 29, 663–678 (1995). https://doi.org/10.1007/BF00041157
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DOI: https://doi.org/10.1007/BF00041157