Skip to main content
SpringerLink
Log in

The Arabidopsis thaliana dhdps gene encoding dihydrodipicolinate synthase, key enzyme of lysine biosynthesis, is expressed in a cell-specific manner

  • Published:
Plant Molecular Biology Aims and scope Submit manuscript

Abstract

Lysine synthesis in prokaryotes, some phycomycetes and higher plants starts with the condensation of L-aspartate-β-semialdehyde (L-ASA) and pyruvate into dihydrodipicolinic acid. The enzyme that catalyses this step, dihydrodipicolinate synthase (DHDPS), is inhibited by the end-product lysine and is therefore thought to have a regulatory control on lysine synthesis. We have cloned and sequenced an Arabidopsis thaliana DNA fragment containing 900 bases upstream of the dhdps coding sequence. A transcriptional fusion of this fragment with the β-glucuronidase reporter gene (uidA, Gus) was used to study the transcription properties of this promoter fragment (DS). No lysine-induced repression on transcription could be detected. Expression of DS-Gus activity in transformed Arabidopsis thaliana and Nicotiana tabacum was found to be cell type-specific. In the vegetative parts of the plant, GUS activity was located in meristems and young vasculature of roots, in vasculature of stem and leaves and in the meristems of young shoots. In flowers, high expression was found in the carpels, style, stigma, developing embryos, tapetum of young anthers and pollen. We demonstrated that the Arabidopsis DS promoter can direct its cell type-specific expression in a heterologous plant, Nicotiana tabacum. The importance of transcriptional regulation of the dhdps gene, and in more general genes involved in amino acid biosynthesis, is discussed.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Bichler J, Herrmann R: Analysis of the promoter of the singlecopy genes for plastocyanin and subunit δ of the chloroplast ATP synthase from spinach. Eur J Biochem 190: 415–426 (1990).

    PubMed  Google Scholar 

  2. Bouvier J, Richaud C, Richaud F, Patte J, Stragier P: Nucleotide sequence and expression of the E. coli dapB gene. J Biol Chem 259: 14829–14834(1984).

    PubMed  Google Scholar 

  3. Boy E, Richaud C, Patte J: Multiple regulation of DAPdecarboxylase synthesis in E. coli K-12. FEMS Microbiol Lett 5: 287–290(1979).

    Google Scholar 

  4. Brears T, Walker E, Coruzzi G: A promoter sequence involved in cell-specific expression of the pea glutamine synthetase GS3A gene in organs of transgenic tobacco and alfalfa. Plant J 1: 235–244(1991).

    PubMed  Google Scholar 

  5. Cattoir-Reynaerts A, Degryse E, Negrutiu I, Jacobs M: Effects of aspartate-derived amino acids on growth of barley and Arabidopsis plants and callus. Z Pflanzenphysiol 101: 67–74 (1981).

    Google Scholar 

  6. Dellaporta S, Wood J, Hicks J: A plant DNA mini preparation: version II. Plant Mol Biol Rep 1: 19–21(1983).

    Google Scholar 

  7. Dereppe C, Bold G, Ghisalba O, Ebert E, Schar H-P: Purification and characterization of dihydrodipicolinate synthase from pea. Plant Physiol 98:813–821(1992).

    Google Scholar 

  8. Feldbrügge M, Sprenger M, Dinkelbach M, Yazaki K, Weisshaar B: Functional analysis of a light-responsive plant bZIP transcriptional regulator. Plant Cell 6: 1607–1621(1994).

    PubMed  Google Scholar 

  9. Flieger K, Tyagi A, Sopory S, Cseplö A, Herrmann R, Oelmüller R: A 42 bp promoter fragment of the gene for subunit III of photosystem I (psaF) is crucial for its activity. Plant J 4: 9–17(1993).

    PubMed  Google Scholar 

  10. Foster R, Gasch A, Kay S, Chua N-H: Analysis of protein/ DNA interactions. In: Koncz C, Chua N-H, Schell J (eds) Methods in Arabidopsis Research, pp. 378–392. World Scientific Publishing, Singapore (1992).

    Google Scholar 

  11. Foster R, Izawa T, Chua N-H: Plant bZIP proteins gather at ACGT elements. FASEB J 8: 192–200(1994).

    PubMed  Google Scholar 

  12. Frankard V, Ghislain M, Jacobs M: Two feedback-insensitive enzymes of the aspartate pathway in Nicotiana sylvestris. Plant Physiol 99: 1285–1293(1992).

    Google Scholar 

  13. Frisch D, Gengenbach B, Tommey A, Sellner J, Somers D, Myers D: Isolation and characterization of dihydrodipicolinate synthase from maize. Plant Physiol 96: 444–452(1991).

    Google Scholar 

  14. Galili G: Regulation of lysine and threonine synthesis. Plant Cell 7: 899–906(1995).

    PubMed  Google Scholar 

  15. Ghislain M, Frankard V, Jacobs M: Dihydrodipicolinate synthase of Nicotiana sylvestris, a chloroplast-localized enzyme of the lysine pathway. Planta 180: 480–486(1990).

    Google Scholar 

  16. Ghislain M, Frankard V, Jacobs M: A dinucleotide mutation in dihydrodipicolinate synthase of Nicotiana sylvestris leads to lysine overproduction. Plant J 8: 733–743(1995).

    PubMed  Google Scholar 

  17. Gilmartin P, Sarokin L, Memelink J, Chua N-H: Molecular light switches for plant genes. Plant Cell 2: 369–378(1990).

    Article  PubMed  Google Scholar 

  18. Green P, Yong M-H, Cuozzo M, Kano-Murakami Y, Silverstein P, Chua N-H: Binding site requirements for pea nuclear protein factor GT-1 correlate with sequences required for light-dependent transcriptional activation of the rbcS-3A gene. EMBO J 7: 4035–4044(1988).

    PubMed  Google Scholar 

  19. Guyer D, Patton D, Ward E: Evidence for cross-pathway regulation of metabolic gene expression in plants. Proc Natl Acad Sci USA 92: 4997–5000(1995).

    PubMed  Google Scholar 

  20. Hérouart D, Van Montagu M, Inzé D: Developmental and environmental regulation of the Nicotiana plumbaginifolia cytosolic Cu/Zn-superoxide dismutase promoter in transgenic tobacco. Plant Physiol 104: 873–880(1994).

    PubMed  Google Scholar 

  21. Hinnebusch A: Mechanisms of gene regulation in the general control of amino acid biosynthesis in Saccharomyces cerevisiae. Microbiol Rev 52: 248–273(1988).

    PubMed  Google Scholar 

  22. Horsch R, Fry J, Hoffman N, Eichholtz D, Rogers S, Fraley R: A simple and general method for transferring genes into plants. Science 277: 1229–1231(1985).

    Google Scholar 

  23. Jefferson R: Assaying chimeric genes in plants: the Gus-gene fusion system. Plant Mol Biol Rep 5: 387–405(1987).

    Google Scholar 

  24. Kumpaisal R, Hashimoto T, Yamada Y: Purification and characterization of dihydrodipicolinate synthase from wheat suspension cultures. Plant Physiol 85: 145–151(1987).

    Google Scholar 

  25. Laber B, Gomis-Rüth F, Romã o M, Huber R: Escherichia coli dihydrodipicolinate synthase. Biochem J 288: 691–695 (1992).

    PubMed  Google Scholar 

  26. Lam E, Chua N-H: GT-1 binding site confers light responsive expression in transgenic tobacco. Science 248: 471–248 (1990).

    PubMed  Google Scholar 

  27. LéJohn H: Enzyme regulation, lysine pathways and cell wall structures as indicators of major lines in evolution in fungi. Nature 231: 164–168(1971).

    PubMed  Google Scholar 

  28. Mazelis M, Whatley F, Whatley J: The enzymology of lysine biosynthesis in higher plants; the occurrence, characterization and some regulatory properties of dihydrodipicolinate synthase. FEBS Lett 84: 236–240(1977).

    PubMed  Google Scholar 

  29. McGrath R, Coruzzi G: A gene network controlling glutamine and asparagine biosynthesis in plants. Plant J 1: 275–280 (1991).

    Article  PubMed  Google Scholar 

  30. Nagy J, Maliga P: Callus induction and plant regeneration from mesophyll protoplasts of Nicotiana sylvestris. Z Pflanzenphysiol 78: 453–455(1976).

    Google Scholar 

  31. Nap P, van Spanje M, DirkseW, Baarda G, Mlynarova L, Loonen A, Grondhuis P, Stiekema W: Activity of the promoter of the Lhca.St.1 gene, encoding the potato apoprotein 2 of the light-harvesting complex of Photosytem I, in transgenic potato and tobacco plants. Plant Mol Biol 23: 605–612(1993).

    PubMed  Google Scholar 

  32. Negrutiu I, Cattoir-Reynaerts A, Verbruggen I, Jacobs M: Lysine overproducer mutants with an altered dihydrodipicolinate synthase from protoplast culture of Nicotiana sylvestris (Spegazzinni and Comes). Theor Appl Genet 68: 11–20 (1984).

    Article  Google Scholar 

  33. Oostindië r-Braaksma F, Feenstra W: Isolation and characterization of chlorate-resistant mutants of Arabidopsis thaliana. Mut Res 19: 175–185(1973).

    Google Scholar 

  34. Peleman J, Boerjan W, Engler G, Seurinck J, Botterman J, Alliotte T, Van Montagu M, Inzé D: Strong cellular preference in the expression of a housekeeping gene of Arabidopsis thaliana encoding S-adenosylmethionine synthetase. Plant Cell 1: 81–93(1989).

    PubMed  Google Scholar 

  35. Ramos F, Dubois E, Piérard A: Control of enzyme synthesis in the lysine biosynthetic pathway of Saccharomyces cerevisiae. Evidence for a regulatory role of gene LYS14. Eur J Biochem 171: 171–176(1988).

    PubMed  Google Scholar 

  36. Richaud C, Richaud F, Martin C, Haziza C, Patte J: Regulation of expression and nucleotide sequence of the E. coli dapD gene. J Biol Chem 259: 14824–14828(1986).

    Google Scholar 

  37. Sambrook J, Fritsch E, Maniatis T: Molecular Cloning: A LaboratoryManual, 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1989).

    Google Scholar 

  38. Shimoni J, Lev-Yadun S, Mattthews B, Galili G: Expression of an aspartate kinase homoserine dehydrogenase gene is subject to specific spatial and temporal regulation in vegetative tissues, flowers and developing seeds. Plant Physiol 113: 695–706 (1997).

    PubMed  Google Scholar 

  39. Stragier P, Richaud F, Borne F, Patte J: Regulation of diaminopimelate decarboxylase synthesis in E. coli. I. Identifi-cation of a lysR gene encoding an activator of the lysA gene. J Mol Biol 168: 307–320(1983).

    PubMed  Google Scholar 

  40. Stragier P, Danos O, Patte J: Regulation of diaminopimelate decarboxylase synthesis in E. coli. II. Nucleotide sequence of the lysA gene and its regulatory region. J Mol Biol 168: 321–331 (1983).

    PubMed  Google Scholar 

  41. Urrestarazu L, Borell C, Bhattacharjee J: General and specific controls of lysine biosynthesis in Saccharomyces cerevisiae. Curr Genet 9: 341–344(1985).

    PubMed  Google Scholar 

  42. Valvekens D, Van Montagu M, Van Lijsebettens M: Agrobacterium tumefaciens-mediated transformation of Arabidopsis thaliana root explants using kanamycin selection. Proc Natl Acad Sci USA 85: 5536–5540(1988).

    Google Scholar 

  43. Vauterin M, Jacobs M: Isolation of a poplar and an Arabidopsis thaliana dihydrodipicolinate synthase cDNA clone. Plant Mol Biol 25: 545–550(1994).

    PubMed  Google Scholar 

  44. Verwoerd C, Dekker B, Hoekema A: A small-scale procedure for rapid isolation of plant RNAs. Nucl Acids Res 17: 2362 (1989).

    PubMed  Google Scholar 

  45. Wallsgrove R, Mazelis M: Spinach leaf dihydrodipicolinate synthase: partial purification and characterization. Phytochemistry 20: 2651–2655(1981).

    Google Scholar 

  46. Wittenbach V, Teaney P, Hanna W, Rayer D, Schloss J: Herbicidal activity of an isopropylmalate dehydrogenase inhibitor. Plant Physiol 106: 321–328(1994).

    PubMed  Google Scholar 

  47. Yang N, Russell D: Maize sucrose synthase-1 promoter directs phloem-specific expression of Gus-gene in transgenic tobacco plants. Proc Natl Acad Sci USA 87: 4144–4148(1990).

    PubMed  Google Scholar 

  48. Yeh P, Sicard A, Sinskey A: General organization of the genes specifically involved in the diaminopimelate-lysine biosynthesic pathway of Corynebacterium glutamicum. Mol Gen Genet 212: 105–111(1988).

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Laboratorium voor Plantengenetica, Vrije Universiteit Brussel, Paardenstraat 65, B-1640, Sint-Genesius Rode, Belgium

    Marc Vauterin, Valérie Frankard & Michel Jacobs

Authors
  1. Marc Vauterin
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Valérie Frankard
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Michel Jacobs
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Vauterin, M., Frankard, V. & Jacobs, M. The Arabidopsis thaliana dhdps gene encoding dihydrodipicolinate synthase, key enzyme of lysine biosynthesis, is expressed in a cell-specific manner. Plant Mol Biol 39, 695–708 (1999). https://doi.org/10.1023/A:1006132428623

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1006132428623

Search

Navigation