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Molecular characterization of a gene for alanine aminotransferase from rice (Oryza sativa)

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Abstract

A cDNA clone encoding alanine aminotransferase (AlaAT) has isolated from randomly sequenced clones derived from a cDNA library of maturing rice seeds by comparison to previously identified genes. The deduced amino acid sequence was 88% and 91% homologous to those of the enzymes from barley and broomcorn millet (Panicum miliaceum), respectively. Using this cDNA as a probe, we isolated and sequenced the corresponding genomic clone. Comparison of the sequences of the cDNA and the genomic gene revealed that the coding region of the gene was interrupted by 14 introns 66 to 1547 bp long. Northern and western blotting analyses showed that the gene was expressed at high levels in developing seeds. When the 5′-flanking region between −930 and +85 from the site of initiation of transcription was fused to a reporter gene for β-glucuronidase (GUS) and then introduced into the rice genome, histochemical staining revealed strong GUS activity in the inner endosperm tissue of developing seeds and weak activity in root tips. Similar tissue-specific expression was also detected by in situ hybridization. These results suggest that AlaAT is involved in nitrogen metabolism during the maturation of rice seed.

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References

  1. Dickinson CD, Evans RP, Nielsen NC: RY repeats are conserved in the 50-flanking region of legume seed protein genes. Nucl Acids Res 16: 371 (1988).

    Google Scholar 

  2. Furuya M: Phytochromes: their molecular species, gene families, and functions. Annu Rev Plant Physiol Plant Mol Biol 44: 617–645 (1993).

    Google Scholar 

  3. Hanley BA, Schuler MA: Plant intron sequences: evidence for distinct groups of introns. Nucl Acids Res 16: 7159–7176 (1988).

    Google Scholar 

  4. Hayakawa T, Yamaya T, Mae T, Ojima K: Changes in the content of two glutamate synthase proteins in spikekets of rice (Oryza sativa) plant during ripening. Plant Physiol 101: 1257–1262 (1993).

    Google Scholar 

  5. Hayakawa T, Nakamura T, Hattori F, Mae T, Ojima K, Yamaya T: Cellular localization of NADH-dependent glutamatesynthase protein in vascular bundles of unexpanded leaf blades and young grains of rice plants. Planta 193: 455–460 (1994).

    Google Scholar 

  6. Hayashi H, Chino M: Chemical composition of phloem sap from the uppermost internode of the rice plants. Plant Cell Physiol 31: 247–251 (1990).

    Google Scholar 

  7. Joshi CP, Zhou H, Huang X, Chiang VL: Context sequences of translation initiation codon in plants. Plant Mol Biol 35: 993–1001 (1997).

    Google Scholar 

  8. Joshi CP: Putative polyadenylation signals in nuclear genes of higher plants: a compilation and analysis. Nucl Acids Res 15: 9627–9640 (1987).

    Google Scholar 

  9. Ishiguro M, Takio K, Suzuki M, Oyama R, Matsuzawa T, Titani K: Complete amino acid sequence of human liver cytosolic alanine aminotransferase (GPT) determined by a combination of conventional and mass spectral methods. Biochemistry 30: 10451–10457 (1991).

    Google Scholar 

  10. Ishiguro M, Suzuki M, Takio K, Matsuzawa T, Titani K: Complete amino acid sequence of rat liver cytosolic alanine aminotransferase. Biochemistry 30: 6048–6053 (1991).

    Google Scholar 

  11. Kamachi K, Yamaya T, Mae T, Ojima K: A role for glutamine synthetase in the emobilization of leaf nitrogen during natural senescence in rice leaves. Plant Physiol 96: 411–417 (1991).

    Google Scholar 

  12. Kawasaki T, Mizuno K, Baba T, Shimada H: Molecular analysis of the gene encoding a rice starch branching enzyme. Mol Gen Genet 237: 10–16 (1998).

    Google Scholar 

  13. Kodama H, Akagi H, Kusumi K, Fujimura T, Iba K: Structure, chromosomal location and expression of a rice gene encoding the microsome !-3 fatty acid desaturase. Plant Mol Biol 33: 493–502 (1997).

    Google Scholar 

  14. Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685 (1970).

    Google Scholar 

  15. Liu J, Hara C, Umeda M, Zhao Y, Okita TW, Uchimiya H: Analysis of randomly isolated cDNAs from developing endosperm of rice (Oryza sativa L.): evaluation of expressed sequence tags, and expression levels of mRNAs. Plant Mol Biol 29: 685–689 (1995).

    Google Scholar 

  16. Masumura T, Hibino T, Kidzu K, Mitsukawa N, Tanaka K, Fijii S: Cloning and characterization of a cDNA encoding a rice 13 kDa prolamin. Mol Gen Genet 221: 1–7 (1990).

    Google Scholar 

  17. Mizuno K, Kawakami, Shimada H, Satoh, Kobayashi E, Okamura S, Arai Y, Baba T: Alteration of the structural properties of starch components by the lack of an isoform of starch branching in rice seeds. J Biol Chem 268: 19084–19091 (1993).

    Google Scholar 

  18. Muench DG, Good AG: Hypoxically inducible barley alanine aminotransferase: cDNA cloning and expression analysis. Plant Mol Biol 24: 417–427 (1994).

    Google Scholar 

  19. Müller M, Knudsen S: The nitrogen response of a barley C-hordein promoter is controlled by positive and negative regulation of the GCN4 and endosperm box. Plant J 4: 343–355 (1993).

    Google Scholar 

  20. Murray MG, Thompson WF: Rapid isolation of highmolecular-weight DNA. Nucl Acids Res 8: 4321–4325 (1980).

    Google Scholar 

  21. Okita TW, Hwang YS, Hnilo J, Kim WT, Aryan AP, Larsen R, Kirishnan HB: Structure and expression of the rice glutelin multigene family. J Biol Chem 264: 12573–12581 (1989).

    Google Scholar 

  22. Panaud O, Chen X, Mcouch SR: Frequency of microsatellite sequences in rice (Oryza sativa L.). Genome 38: 1170–1176 (1995).

    Google Scholar 

  23. Sambrook J, Fritsch EF, Maniatis T: Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY (1989).

    Google Scholar 

  24. Sasaki T, Song J, Koga-Ban Y, Matsui E, Fang F, Higo H, Nagasaki H, Hori M, Miya M, Maruyama-Kayano E, Takiguchi T, Takasuga A, Niki T, Ishimaru K, Ikeda H, Yamamoto Y, Murai Y, Ohta I, Miyadera N, Havakkala I, Minobe Y: Toward cataloguing all rice genes: large-scale sequencing of randomly chosen rice cDNAs from a callus cDNA library. Plant J 6: 615–624 (1994)

    Google Scholar 

  25. Shen B, Carneiro N, Torres-Jerez I, Stevenson B, McCreery T, Helentjaris T, Baysdorfer C, Almira E, Ferl R, Habben JE, Larkins B: Partial sequencing and mapping of clones from two maize cDNA libralies. Plant Mol Biol 26: 1085–1101 (1994).

    Google Scholar 

  26. Son D, Sugiyama T: Molecular cloning of an alanine aminotransferase from NAD-malic enzyme type C4 plant Panicum miliaceum. Plant Mol Biol 20: 705–713 (1992).

    Google Scholar 

  27. 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).

    Google Scholar 

  28. Steinkellner H, Fluch S, Turetschek E, Lexer C, Streiff R, Kremer A, Burg K, Gloss: Identification and characterization of (GA/CT)n-microsatellite loci from Quercus petraea. Plant Mol Biol 33: 1093–1096 (1997).

    Google Scholar 

  29. Takaiwa F, Oono K, Sugiura M: The complete nucleotide sequence of a rice 17S rRNA gene. Nucl Acids Res 12: 5441–5448 (1984).

    Google Scholar 

  30. Takaiwa F, Kikuchi S, Oono K: A rice glutelin family-a major type of glutelin mRNAs can be divided into two classes. Mol Gen Genet 208: 15–22 (1987).

    Google Scholar 

  31. Takaiwa F, Oono K: Genomic DNA sequences of two new genes for new storage protein glutelin in rice. Jpn J Genet 66: 161–171 (1991).

    Google Scholar 

  32. Takaiwa F, Oono K, Wing D, Kato A: Sequence of three members and expression of a new major subfamily of glutelin genes from rice. Plant Mol Biol 17: 875–885 (1991).

    Google Scholar 

  33. Takaiwa F, Yamanouchi U, Yoshihara T, Washida H, Tanabe F, Kato K, Yamada K: Characterization of common cisregulatory elements responsible for the endosperm-specific expression of members of the rice glutelin multigene family. Plant Mol Biol 30: 1207–1221 (1996).

    Google Scholar 

  34. Tanase S, Kojima H, Morino Y: Pyridoxal 50-phosphate binding site of pig heart alanine aminotransferase. Biochemistry 18: 3002–3007 (1979).

    Google Scholar 

  35. Toki S, Takamatsu S, Nojiri C, Ooba S, Anzai H, Iwata M, Christensen AH, Quail PH, Uchimiya H: Expression of a maize ubiquitin gene promoter-bar chimeric gene in transgenic rice plants. Plant Physiol 100: 1503–1507 (1992).

    Google Scholar 

  36. Yoshihara T, Washida H, Takaiwa F: A 45-bp proximal region containing AACA and GCN4 motif is sufficient to confer endosperm-specific expression of the rice storage protein glutelin gene, GluA-3. FEBS Lett 383: 213–218 (1996).

    Google Scholar 

  37. Wu C-Y, Suzuki A, Washida H, Takaiwa F: The GCN4 motif in a rice glutelin gene is essential for endosperm-specific gene expression and is activated by Opaque-2 in transgenic rice plants. Plant J 14: 673–684 (1998).

    Google Scholar 

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Authors and Affiliations

  1. Department of Biotechnology, National Institute of Agrobiological Resources, Tsukuba, Ibaraki, 305-8602, Japan

    Hidehiko Kikuchi, Sakiko Hirose, Seiichi Toki & Fumio Takaiwa

  2. Department of Biological Science, Shimane University, Matsue, 690, Japan

    Kazuhito Akama

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  1. Hidehiko Kikuchi
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  2. Sakiko Hirose
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  3. Seiichi Toki
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  4. Kazuhito Akama
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  5. Fumio Takaiwa
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Kikuchi, H., Hirose, S., Toki, S. et al. Molecular characterization of a gene for alanine aminotransferase from rice (Oryza sativa). Plant Mol Biol 39, 149–159 (1999). https://doi.org/10.1023/A:1006156214716

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