Abstract
A 1.75 kb cDNA containing the entire coding sequence of the hypoxically inducible alanine aminotransferase (AlaAT) from barley roots was isolated and sequenced. This clone has an open reading frame of 1446 bp, and a deduced amino acid sequence of 482 residues, giving an estimated protein molecular mass of 52 885 Da. RNA blot analysis of barley root tissue showed a 4-fold increase of a single AlaAT-2 mRNA band after 12–24 hours of hypoxic stress, followed by a decrease in message levels after 48 h of hypoxic conditions. AlaAT-2 protein concentration increased in a similar pattern to AlaAT activity in root tissue, to almost 6-fold the aerobic level after 96 h of hypoxic stress. AlaAT-2 activity increased more than 2-fold in roots of Panicum miliaceum exposed to hypoxia, and is the same isoform as the light inducible AlaAT in P. miliaceum leaves. The unique expression patterns of AlaAT-2 in root and leaf tissue upon exposure to different environmental stimuli is also discussed.
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References
Alexander DC, McKnight TD, Williams BG: A simplified and efficient vector-primer cDNA cloning system. Gene 31: 79–89 (1984).
Davies DD: Anaerobic metabolism and the production of organic acids. In: DD Davies (ed) The Biochemistry of Plants, vol. 2, pp 581–611. Academic Press New York (1980).
Dennis ES, Gerlach WL, Walker JC, Lavin M, Peacock WJ: Anaerobically regulated aldolase gene in maize, a chimeric origin? J Mol Biol 202: 759–767 (1988).
Effer WR, Ranson SL: Some effects of oxygen concentration on levels of respiratory intermediates in buckwheat seedlings. Plant Physiol 42: 1053–1058 (1967).
Gerlach WL, Pryor AJ, Dennis ES, Ferl RJ, Sachs MM, Peacock WJ: cDNA cloning and induction of the alcohol dehydrogenase gene (Adh1) of maize. Proc Natl Acad Sci USA 79: 2981–2985 (1982).
Givan CV: Aminotransferases in higher plants. In: Stupf PK, Conn EE (eds) The Biochemistry of Plants. A Comprehensive Treatise, pp 346–350. Academic Press, New York (1980).
Good AG, Crosby WL: Induction of alcohol dehydrogenase and lactate dehydrogenase in hypoxically induced barley. Plant Physiol 90: 860–866 (1989).
Good AG, Crosby WL: Anaerobic induction of alanine aminotransferase in barley root tissue. Plant Physiol 90: 1305–1309 (1989).
Good AG, Muench DG: Purification and characterization of an anaerobically induced alanine aminotransferase from barley roots. Plant Physiol 99: 1520–1525 (1992).
Good AG, Muench DG: Long-term anaerobic metabolism in root tissue, metabolic products of pyruvate metabolism. Plant Physiol 101: 1163–1168 (1993).
Hatch MD: C4 photosynthesis: a unique blend of modified biochemistry, anatomy and ultrastructure. Biochim Biophys Acta 895: 6048–6053 (1987).
Ho T-HD, Sachs MM: Stress induced proteins: characterization and the regulation of their synthesis. In: Stumpf PK, Conn EE (eds) The Biochemistry of Plants: A Comprehensive Treatise, vol. 15 pp 362–367. Academic Press (1989).
Hondred D, Hanson AD: Hypoxically inducible barley lactate dehydrogenase: cDNA cloning and molecular analysis. Proc Natl Acad Sci USA 87: 7300–7304 (1990).
Hoffman NE, Bent AF, Hanson AD: Induction of lactate dehydrogenase isozymes by oxygen deficit in barley root tissue. Plant Physiol 82: 658–663 (1986).
Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685 (1970).
Lal SK, Johnson S, Conway T, Kelley PM: Characterization of a maize cDNA that complements an enolase-deficient mutant of Escherichia coli. Plant Mol Biol 16: 787–795 (1991).
Muench DG, Archibold OW, Good AG: Hypoxic metabolism in wild rice (Zizania palustris L.): enzyme induction and metabolite production. Physiol Plant, in press (1993).
Olive MR, Walker JC, Singh K, Ellis JG, Llewellyn D, Peacock WJ, Dennis ES: The anaerobic response element. In: Herman RG, Larkins BA (eds) Plant Molecular Biology vol. 2, pp. 673–674. Plenum Press, New York (1991).
Russell DA, Sachs MM: Differential expression and sequence analysis of the maize glyceraldehyde-3-phosphate dehydrogenase gene family. Plant Cell 1: 793–803 (1989).
Sachs MM, Freeling M, Okimoto R: The anaerobic proteins of maize. Cell 20: 761–767 (1980).
Sanger F, Nicklen S, Coulson AR: DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74: 5463–5467 (1977).
Selden RF: Analysis of RNA by northern hybridization. In: Ausubel FM, Brent R, Kingston Re, Moore DD, Seidman JG, Smith JA, Struhl K (eds) Current Protocols in Molecular Biology, pp 4.9.1–4.9.8. John Wiley, New York (1989).
Smith AM, ap Rees T: Effects of anaerobiosis on carbohydrate oxidation by roots of Pisum sativum. Phytochemistry 18: 1453–1458 (1979).
Son D, Jo J, Sugiyama T: Purification and characterization of alanine aminotransferase from Panicum miliaceum leaves. Arch Biochem Biophys 289: 262–266 (1991).
Son D, Kobe A, Sugiyama T: Nitrogen-dependent regulation of the gene for alanine aminotransferase which is involved in the C4 pathway of Panicum miliaceum. Plant Cell Physiol 33: 507–509 (1992).
Son D, Sugiyama T: Molecular cloning of alanine aminotransferase from NAD-malic enzyme type C4 plant Panicum miliaceum. Plant Mol Biol 20: 705–713 (1992).
Streeter JG, Thompson JF: Anaerobic accumulation of γ-aminobutyrate and alanine in radish leaves (Raphinus sativus). Plant Physiol 49: 572–578 (1972).
Watson NR, Peschke VM, Russel DA, Sachs MM: Analysis of the L-alanine:2-oxoglutarate aminotransferase isozymes in maize. Biochem Genet 30: 371–383 (1992).
Yeh K-W, Juang R-H, Su J-C: A rapid and efficient method for RNA isolation from plants with high carbohydrate content. Focus 13: 102–103 (1990).
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Muench, D.G., Good, A.G. Hypoxically inducible barley alanine aminotransferase: cDNA cloning and expression analysis. Plant Mol Biol 24, 417–427 (1994). https://doi.org/10.1007/BF00024110
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DOI: https://doi.org/10.1007/BF00024110