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. 1990 Feb;9(2):323–332. doi: 10.1002/j.1460-2075.1990.tb08114.x

Dark-induced and organ-specific expression of two asparagine synthetase genes in Pisum sativum.

F Y Tsai 1, G M Coruzzi 1
PMCID: PMC551669  PMID: 1968003

Abstract

Nucleotide sequence analysis of cDNAs for asparagine synthetase (AS) of Pisum sativum has uncovered two distinct AS mRNAs (AS1 and AS2) encoding polypeptides that are highly homologous to the human AS enzyme. The amino-terminal residues of both AS1 and AS2 polypeptides are identical to the glutamine-binding domain of the human AS enzyme, indicating that the full-length AS1 and AS2 cDNAs encode glutamine-dependent AS enzymes. Analysis of nuclear DNA shows that AS1 and AS2 are each encoded by single genes in P.sativum. Gene-specific Northern blot analysis reveals that dark treatment induces high-level accumulation of AS1 mRNA in leaves, while light treatment represses this effect as much as 30-fold. Moreover, the dark-induced accumulation of AS1 mRNA was shown to be a phytochrome-mediated response. Both AS1 and AS2 mRNAs also accumulate to high levels in cotyledons of germinating seedlings and in nitrogen-fixing root nodules. These patterns of AS gene expression correlate well with the physiological role of asparagine as a nitrogen transport amino acid during plant development.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Andrulis I. L., Chen J., Ray P. N. Isolation of human cDNAs for asparagine synthetase and expression in Jensen rat sarcoma cells. Mol Cell Biol. 1987 Jul;7(7):2435–2443. doi: 10.1128/mcb.7.7.2435. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Back E., Burkhart W., Moyer M., Privalle L., Rothstein S. Isolation of cDNA clones coding for spinach nitrite reductase: complete sequence and nitrate induction. Mol Gen Genet. 1988 Apr;212(1):20–26. doi: 10.1007/BF00322440. [DOI] [PubMed] [Google Scholar]
  3. Biggin M. D., Gibson T. J., Hong G. F. Buffer gradient gels and 35S label as an aid to rapid DNA sequence determination. Proc Natl Acad Sci U S A. 1983 Jul;80(13):3963–3965. doi: 10.1073/pnas.80.13.3963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Boutry M., Chua N. H. A nuclear gene encoding the beta subunit of the mitochondrial ATP synthase in Nicotiana plumbaginifolia. EMBO J. 1985 Sep;4(9):2159–2165. doi: 10.1002/j.1460-2075.1985.tb03910.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Capdevila A. M., Dure L. Developmental Biochemistry of Cottonseed Embryogenesis and Germination: VIII. Free Amino Acid Pool Composition during Cotyledon Development. Plant Physiol. 1977 Feb;59(2):268–273. doi: 10.1104/pp.59.2.268. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Crawford N. M., Campbell W. H., Davis R. W. Nitrate reductase from squash: cDNA cloning and nitrate regulation. Proc Natl Acad Sci U S A. 1986 Nov;83(21):8073–8076. doi: 10.1073/pnas.83.21.8073. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cullimore J. V., Gebhardt C., Saarelainen R., Miflin B. J., Idler K. B., Barker R. F. Glutamine synthetase of Phaseolus vulgaris L.: organ-specific expression of a multigene family. J Mol Appl Genet. 1984;2(6):589–599. [PubMed] [Google Scholar]
  8. Dilworth M. F., Dure L. Developmental biochemistry of cotton seed embryogenesis and germination: x. Nitrogen flow from arginine to asparagine in germination. Plant Physiol. 1978 Apr;61(4):698–702. doi: 10.1104/pp.61.4.698. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Felton J., Michaelis S., Wright A. Mutations in two unlinked genes are required to produce asparagine auxotrophy in Escherichia coli. J Bacteriol. 1980 Apr;142(1):221–228. doi: 10.1128/jb.142.1.221-228.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Huber T. A., Streeter J. G. Asparagine biosynthesis in soybean nodules. Plant Physiol. 1984 Mar;74(3):605–610. doi: 10.1104/pp.74.3.605. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Humbert R., Simoni R. D. Genetic and biomedical studies demonstrating a second gene coding for asparagine synthetase in Escherichia coli. J Bacteriol. 1980 Apr;142(1):212–220. doi: 10.1128/jb.142.1.212-220.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Jones G. E. L-Asparagine auxotrophs of Saccharomyces cerevisiae: genetic and phenotypic characterization. J Bacteriol. 1978 Apr;134(1):200–207. doi: 10.1128/jb.134.1.200-207.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Joy K. W., Ireland R. J., Lea P. J. Asparagine synthesis in pea leaves, and the occurrence of an asparagine synthetase inhibitor. Plant Physiol. 1983 Sep;73(1):165–168. doi: 10.1104/pp.73.1.165. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kay S. A., Keith B., Shinozaki K., Chye M. L., Chua N. H. The rice phytochrome gene: structure, autoregulated expression, and binding of GT-1 to a conserved site in the 5' upstream region. Plant Cell. 1989 Mar;1(3):351–360. doi: 10.1105/tpc.1.3.351. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kern R., Chrispeels M. J. Influence of the axis on the enzymes of protein and amide metabolism in the cotyledons of mung bean seedlings. Plant Physiol. 1978 Nov;62(5):815–819. doi: 10.1104/pp.62.5.815. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lissemore J. L., Quail P. H. Rapid transcriptional regulation by phytochrome of the genes for phytochrome and chlorophyll a/b-binding protein in Avena sativa. Mol Cell Biol. 1988 Nov;8(11):4840–4850. doi: 10.1128/mcb.8.11.4840. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Loh E. Y., Elliott J. F., Cwirla S., Lanier L. L., Davis M. M. Polymerase chain reaction with single-sided specificity: analysis of T cell receptor delta chain. Science. 1989 Jan 13;243(4888):217–220. doi: 10.1126/science.2463672. [DOI] [PubMed] [Google Scholar]
  18. Lycett G. W., Croy R. R., Shirsat A. H., Richards D. M., Boulter D. The 5'-flanking regions of three pea legumin genes: comparison of the DNA sequences. Nucleic Acids Res. 1985 Sep 25;13(18):6733–6743. doi: 10.1093/nar/13.18.6733. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Mösinger E., Batschauer A., Schäfer E., Apel K. Phytochrome control of in vitro transcription of specific genes in isolated nuclei from barley (Hordeum vulgare). Eur J Biochem. 1985 Feb 15;147(1):137–142. doi: 10.1111/j.1432-1033.1985.tb08729.x. [DOI] [PubMed] [Google Scholar]
  20. Nakamura M., Yamada M., Hirota Y., Sugimoto K., Oka A., Takanami M. Nucleotide sequence of the asnA gene coding for asparagine synthetase of E. coli K-12. Nucleic Acids Res. 1981 Sep 25;9(18):4669–4676. doi: 10.1093/nar/9.18.4669. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Pearson W. R., Lipman D. J. Improved tools for biological sequence comparison. Proc Natl Acad Sci U S A. 1988 Apr;85(8):2444–2448. doi: 10.1073/pnas.85.8.2444. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Ramos F., Wiame J. M. Two asparagine synthetases in Saccharomyces cerevisiae. Eur J Biochem. 1980 Jul;108(2):373–377. doi: 10.1111/j.1432-1033.1980.tb04732.x. [DOI] [PubMed] [Google Scholar]
  23. Streeter J. G. Asparaginase and asparagine transaminase in soybean leaves and root nodules. Plant Physiol. 1977 Aug;60(2):235–239. doi: 10.1104/pp.60.2.235. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Tingey S. V., Tsai F. Y., Edwards J. W., Walker E. L., Coruzzi G. M. Chloroplast and cytosolic glutamine synthetase are encoded by homologous nuclear genes which are differentially expressed in vivo. J Biol Chem. 1988 Jul 15;263(20):9651–9657. [PubMed] [Google Scholar]
  25. Tingey S. V., Walker E. L., Coruzzi G. M. Glutamine synthetase genes of pea encode distinct polypeptides which are differentially expressed in leaves, roots and nodules. EMBO J. 1987 Jan;6(1):1–9. doi: 10.1002/j.1460-2075.1987.tb04710.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Urquhart A. A., Joy K. W. Use of Phloem exudate technique in the study of amino Acid transport in pea plants. Plant Physiol. 1981 Sep;68(3):750–754. doi: 10.1104/pp.68.3.750. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Van Heeke G., Schuster S. M. Expression of human asparagine synthetase in Escherichia coli. J Biol Chem. 1989 Apr 5;264(10):5503–5509. [PubMed] [Google Scholar]

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