Abstract
ADP-glucose pyrophosphorylase (AGPase) is one of the major enzymes involved in starch biosynthesis in higher plants. We report here the molecular cloning of two cDNAs encoding so far uncharacterized isoforms (AGP S2 and AGP S3) of the potato enzyme. Sequence analysis shows that the two polypeptides are more homologous to previously identified large subunit polypeptides from potato and other plant species than to small subunit isoforms. This observation suggests that AGP S2 and AGP S3 represent novel large subunit polypeptides. agpS2 is expressed in several tissues of the potato plant, including leaves and tubers. Expression was stronger in sink leaves than in source leaves, indicating developmental regulation. In leaves, agpS2 expression was induced 2- to 3-fold by exogenous sucrose; therefore, agpS2 represents a new sucrose-responsive gene of starch metabolism. Expression of agpS3 was restricted to tubers: no agpS3 expression could be seen in leaves of different developmental stages, or when leaves were incubated in sucrose. Therefore, agpS3 represents the only AGPase gene so far characterized from potato, which is not expressed in leaves. Conversely, all four AGPase isoforms known from potato are expressed in tubers.
Similar content being viewed by others
References
Ainsworth C, Tarvis M, Clark J (1993) Isolation and analysis of a cDNA clone encoding the small subunit of ADP-glucose pyrophosphorylase from wheat. Plant Mol Biol 23:23–33
Amasino RM (1986) Acceleration of nucleic acid hybridisation rate by polyethylene glycol. Anal Biochem 152:304–307
Anderson JM, Hnilo J, Larson R, Okita TW, Morell M, Preiss J (1989) The encoded primary sequence of a rice ADP-glucose pyrophosphorylase subunit and its homology to the bacterial enzyme. J Biol Chem 264:12238–12242
Bae JM, Giroux M, Hannah L (1990) Cloning and characterization of the brittle-2 gene of maize. Maydica 35:317–322
Baecker PA, Furlong CE, Preiss J (1983) Biosynthesis of bacterial glycogen. Primary structure of Escherichia coli ADP-glucose synthetase as deduced from the nucleotide sequence of the glgC gene. J Biol Chem 258:5084–5088
Bhave MR, Lawrence S, Barton C, Hannah LC (1990) Identification and molecular characterization of shrunken-2 cDNA clones of maize. Plant Cell 2:581–588
Devereux J, Haeberli P, Smithies O (1984) A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res 12:387–395
Gebhardt C, Ritter E, Salamini F (1994) RFLP map of the potato. In: Vasil IK, Phillips RL (eds) Advances in cellular and molecular biology of plants. Vol 1: DNA-based markers in plants. Kluwer Academic Publishers, Dordrecht, The Netherlands, pp 271–285
Ghosh HP, Preiss J (1966) Adenosine diphosphate glucose pyrophosphorylase: a regulatory enzyme in the biosynthesis of starch in spinach leaf chloroplasts. J Biol Chem 241:4491–4504
Hylton C, Smith AM (1992) The rb mutation of peas causes structural and regulatory changes in ADP glucose pyrophosphorylase from developing embryos. Plant Physiol 99:1626–1634
Iglesias AA, Barry GF, Meyer C, Bloksberg L, Nakata PA, Greene T, Laughlin MJ, Okita TW Kishore GM, Preiss J (1993) Expression of the potato tuber ADP-glucose pyrophosphorylase in Escherichia coli. J Biol Chem 268:1081–1086
Jardin du P, Berhin A (1991) Isolation and sequence analysis of a cDNA clone encoding a subunit of the ADP-glucose pyrophosphorylase of potato tuber amyloplasts. Plant Mol Biol 16:349–351
Joshi CP (1987) Putative polyadenylation signals in nuclear genes of higher plants: a compilation and analysis. Nucleic Acids Res 15:9627–9640
Kleczkowski LA, Villand P, Lüthi E, Olsen O-A, Preiss J (1993) Insensitivity of barley endosperm ADP-glucose pyrophosphorylase to 3-phosphoglycerate and orthophosphate regulation. Plant Physiol 101:179–186
Koch KE, Nolte KD, Duke ER, McCarty DR, Avigne WT (1992) Sugar levels modulate differential expression of maize sucrose synthase genes. Plant Cell 4: 59–69
Koßmann J, Visser RGF, Mülller-Röber B, Willmitzer L, Sonnewald U (1991) Cloning and expression analysis of a potato cDNA that encodes branching enzyme: evidence for co-expression of starch biosynthetic genes. Mol Gen Genet 230: 39–44
Kumar A, Ghosh P, Lee YM, Hill MA, Preiss J (1989) Biosynthesis of bacterial glycogen. Determination of the amino acid changes that alter the regulatory properties of a mutant Escherichia coli ADP-glucose synthetase. J Biol Chem 264:10464–10471
Lee YM, Preiss J (1986) Covalent modification of substrate-binding sites of Escherichia coli ADP-glucose synthetase. Isolation and structural characterization of 8-azido-ADP-glucose-incorporated peptides. J Biol Chem 261:1058–1064
Lee YM, Kumar A, Preiss J (1987) Amino acid sequence of an Escherichia coli ADPglucose synthetase allosteric mutant as deduced from the DNA sequence of the glgC gene. Nucleic Acids Res 15:10603
Lin TP, Caspar T, Somerville C, Preiss J (1988) Isolation and characterization of a starchless mutant of Arabidopsis thaliana (L.) Heynh. lacking ADPglucose pyrophosphorylase activity. Plant Physiol 86:1131–1135
Logemann J, Schell J, Willmitzer L (1987) Improved method for the isolation of RNA from plant tissues. Anal Biochem 163:21–26
Morell MK, Bloom M, Knowles V, Preiss J (1987) Subunit structure of spinach leaf ADP-glucose pyrophosphorylase. Plant Physiol 85:182–187
Morell M, Bloom M, Preiss J (1988) Affinity labeling of the allosteric activator site(s) of spinach leaf ADP-glucose pyrophosphorylase. J Biol Chem 263:633–637
Müller-Röber B (1992) Untersuchungen zur Struktur and Funktion der ADP-Glucose Pyrophosphorylase aus Kartoffel unter Einsatz transgener Pflanzen. PhD Thesis, Freie Universität Berlin
Müller-Röber B, Koßmann J, Hannah LC, Willmitzer L, Sonnewald U (1990) Only one of two different ADP-glucose pyrophosphorylase genes from potato responds strongly to elevated levels of sucrose. Mol Gen Genet 224:136–146
Müller-Röber B, Sonnewald U, Willmitzer L (1992) Antisense inhibition of the ADP-glucose pyrophosphorylase in transgenic potatoes leads to sugar-storing tubers and influences tuber formation and expression of tuber storage proteins. EMBO J 11:1229–1238
Müller-Röber B, La Cognata U, Sonnewald U, Willmitzer L (1994) A truncated version of an ADP-glucose pyrophosphorylase promoter from potato specifies guard-cell selective expression in transgenic plants. Plant Cell 6:601–612
Nakamura Y, Kawaguchi K (1992) Multiple forms of ADPglucose pyrophosphorylase of rice endosperm. Physiol Plant 84:336–342
Nakata PA, Greene TW, Anderson JM, Smith-White BJ, Okita TW, Preiss J (1991) Comparison of the primary sequences of two potato tuber ADP-glucose pyrophosphorylase subunits. Plant Mol Biol 17:1089–1093
Neuhaus HE, Stitt M (1990) Control analysis of photosynthate partitioning. Impact of reduced activity of ADP-glucose pyrophosphorylase or plastid phosphoglucomutase on the fluxes to starch and sucrose in Arabidopsis thaliana (L.) Heynh. Planta 182:445–454
Okita TW, Nakata PA, Anderson JM, Sowokinos J, Morell M, Preiss J (1990) The subunit structure of potato tuber ADPglucose pyrophosphorylase. Plant Physiol 93:785–790
Olive MR, Ellis RJ, Schuch WW (1989) Isolation and nucleotide sequences of cDNA clones encoding ADP-glucose pyrophosphorylase polypeptides from wheat leaf and endosperm. Plant Mol Biol 12:525–538
Outlaw WC Jr, Tarczynski MC (1984) Guard cell starch biosynthesis regulated by effectors of ADP-glucose pyrophosphorylase. Plant Physiol 74:424–429
Parsons TF, Preiss J (1978a) Biosynthesis of bacterial glycogen: incorporation of pyridoxal phosphate into the allosteric activator site and an ADP-glucose-protected pyridoxal phosphate-binding site of Escherichia coli B ADP-glucose synthetase. J Biol Chem 253:6197–6202
Parsons TF, Preiss J (1978b) Biosynthesis of bacterial glycogen: isolation and characterization of the pyridoxal-P allosteric activator site and the ADP-glucose-protected pyridoxal-P binding site of Escherichia coli B ADP-glucose synthase. J Biol Chem 253:7638–7645
Pearson WR, Lipman DJ (1988) Improved tools for biological sequence comparison. Proc Natl Acad Sci USA 85:2444–2448
Pettersson G, Ryde-Pettersson U (1989) Metabolites controlling the rate of starch synthesis in the chloroplast of C3 plants. Eur J Biochem 179:169–172
Plaxton WC, Preiss J (1987) Purification and properties of nonproteolytically degraded ADPglucose pyrophosphorylase from maize endosperm. Plant Physiol 83:105–112
Preiss J (1988) In: Preiss J (ed) Biosynthesis of starch and its regulation. The biochemistry of plants, vol 14. Academic Press, New York, pp 181–254
Preiss J (1991) Biology and molecular biology of starch synthesis and its regulation. Ox Surv Plant Mol Cell Biol 7:59–114
Robinson NL, Zeiger E, Preiss J (1983) Regulation of ADP-glucose synthesis in guard cells of Commelina communis. Plant Physiol 73:862–864
Rogers SO, Bendich AJ (1985) Extraction of DNA from milligram amounts of fresh, herbarium and mummified plant tissue. Plant Mol Biol 5:69–76
Satanoubat M, Belliard G (1989) The steady-state level of potato sucrose synthase mRNA is dependent on wounding, anaerobiosis and sucrose concentration. Gene 84:181–185
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York
Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
Sanwal GG, Greenberg E, Hardie J, Cameron EC, Preiss J (1968) Regulation of starch biosynthesis in plant leaves: activation and inhibition of ADPglucose pyrophosphorylase. Plant Physiol 43:417–427
Smith-White BJ, Preiss J (1992) Comparison of proteins of ADPglucose pyrophosphorylase from diverse sources. J Mol Evol 34:449–464
Smith-White B, Preiss J (1994) Suggested mnemonics for cloned DNA corresponding to enzymes involved in starch metabolism. Plant Mol Biol Rep 12 (CPGN Suppl) 67–71
Sowokinos JR (1976) Pyrophosphorylases in Solanum tuberosum. I. Changes in ADPglucose and UDPglucose pyrophosphorylase activities associated with starch biosynthesis during tuberization, maturation, and storage of potatoes. Plant Physiol 57:63–68
Sowokinos JR, Preiss J (1982) Pyrophosphorylases in Solanum tuberosum. III. Purification, physical, and catalytic properties of ADPglucose pyrophosphorylase in potatoes. Plant Physiol 69:1459–1466
Stark DM, Timmermann KP, Barry GF, Preiss J, Kishore GM (1992) Regulation of the amount of starch in plant tissues by ADP glucose pyrophosphorylase. Science 258:287–292
Tsai CY, Nelson OE (1966) Starch-deficient maize mutant lacking adenosine diphosphate glucose pyrophosphorylase activity. Science 151:341–343
Villand P, Aalen R, Olsen O-A, Lüthi E, Lönneborg A, Kleczkowski LA (1992a) PCR amplification and sequence of cDNA clones for the small and large subunits of ADP-glucose pyrophosphorylase from barley tissues. Plant Mol Biol 19:381–389
Villand P, Olsen O-A, Kilian A, Kleczkowski LA (1992b) ADPglucose pyrophosphorylase large subunit cDNA from barley endosperm. Plant Physiol 100:1617–1618
Villand P, Olsen O-A, Kleczkowski LA (1993) Molecular characterization of multiple cDNA clones for ADP-glucose pyrophosphorylase from Arabidopsis thaliana. Plant Mol Biol 23:1279–1284
Visser RGF, Stolte A, Jacobsen E (1991) Expression of a chimaeric granule-bound starch synthase-GUS gene in transgenic plants. Plant Mol Biol 17:691–699
Author information
Authors and Affiliations
Additional information
Communicated by H. Saedler
Rights and permissions
About this article
Cite this article
La Cognata, U., Willmitzer, L. & Müller-Röber, B. Molecular cloning and characterization of novel isoforms of potato ADP-glucose pyrophosphorylase. Molec. Gen. Genet. 246, 538–548 (1995). https://doi.org/10.1007/BF00298960
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00298960