Skip to main content
PMC home page
The Plant Cell logoLink to The Plant Cell
. 1993 Aug;5(8):877–886. doi: 10.1105/tpc.5.8.877

A 22-bp fragment of the pea lectin promoter containing essential TGAC-like motifs confers seed-specific gene expression.

S de Pater 1, K Pham 1, N H Chua 1, J Memelink 1, J Kijne 1
PMCID: PMC160323  PMID: 8400870

Abstract

To elucidate the molecular mechanisms responsible for seed-specific gene expression in plants, the promoter of the pea lectin (psl) gene, encoding an abundant seed protein, was used as a model. Leaf and seed nuclear proteins bound to a region in the psl promoter containing three overlapping TGAC-like motifs, which have been shown to be a binding site for basic/leucine zipper proteins, including TGA1a. A trimer of a 22-bp region of the psl promoter, containing the TGAC-like motifs, coupled to a heterologous minimal promoter conferred low reporter gene expression in root, stem, and leaf and high expression in seed of transgenic tobacco. Expression increased during the midmaturation stage of seed development and was observed in the endosperm as well as in the embryo, where it strongly decreased within a few days after germination. This expression pattern is qualitatively identical to the expression pattern conferred by a 2000-bp fragment of the psl promoter. Nucleotides within the TGAC-like motifs important for in vitro binding are also essential for in vivo transcription activation in vegetative tissue as well as in seed. The electrophoretic mobility of a DNA-protein complex containing seed nuclear protein was different from that formed with leaf nuclear protein. Furthermore, the TGA1a steady state mRNA level in immature seed was relatively low. These results suggest that a seed-specific factor different from TGA1a, but with similar binding specificity, is responsible for gene activation in seed. We conclude that the 22-bp region contains all the information, including an essential TGAGTCATCA sequence, necessary for seed-specific expression and very likely plays an essential role in the seed-specific expression pattern of the psl gene.

Full Text

The Full Text of this article is available as a PDF (2.2 MB).

Selected References

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

  1. A simple and general method for transferring genes into plants. Science. 1985 Mar 8;227(4691):1229–1231. doi: 10.1126/science.227.4691.1229. [DOI] [PubMed] [Google Scholar]
  2. Benfey P. N., Ren L., Chua N. H. Tissue-specific expression from CaMV 35S enhancer subdomains in early stages of plant development. EMBO J. 1990 Jun;9(6):1677–1684. doi: 10.1002/j.1460-2075.1990.tb08291.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  4. Chamberland S., Daigle N., Bernier F. The legumin boxes and the 3' part of a soybean beta-conglycinin promoter are involved in seed gene expression in transgenic tobacco plants. Plant Mol Biol. 1992 Sep;19(6):937–949. doi: 10.1007/BF00040526. [DOI] [PubMed] [Google Scholar]
  5. Fromm H., Katagiri F., Chua N. H. The tobacco transcription activator TGA1a binds to a sequence in the 5' upstream region of a gene encoding a TGA1a-related protein. Mol Gen Genet. 1991 Oct;229(2):181–188. doi: 10.1007/BF00272154. [DOI] [PubMed] [Google Scholar]
  6. Gatehouse J. A., Bown D., Evans I. M., Gatehouse L. N., Jobes D., Preston P., Croy R. R. Sequence of the seed lectin gene from pea (Pisum sativum L.). Nucleic Acids Res. 1987 Sep 25;15(18):7642–7642. doi: 10.1093/nar/15.18.7642. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Goldberg R. B., Barker S. J., Perez-Grau L. Regulation of gene expression during plant embryogenesis. Cell. 1989 Jan 27;56(2):149–160. doi: 10.1016/0092-8674(89)90888-x. [DOI] [PubMed] [Google Scholar]
  8. Green P. J., Kay S. A., Chua N. H. Sequence-specific interactions of a pea nuclear factor with light-responsive elements upstream of the rbcS-3A gene. EMBO J. 1987 Sep;6(9):2543–2549. doi: 10.1002/j.1460-2075.1987.tb02542.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hayashi S., Scott M. P. What determines the specificity of action of Drosophila homeodomain proteins? Cell. 1990 Nov 30;63(5):883–894. doi: 10.1016/0092-8674(90)90492-w. [DOI] [PubMed] [Google Scholar]
  10. Higgins T. J., Chrispeels M. J., Chandler P. M., Spencer D. Intracellular sites of synthesis and processing of lectin in developing pea cotyledons. J Biol Chem. 1983 Aug 10;258(15):9550–9552. [PubMed] [Google Scholar]
  11. Izawa T., Foster R., Chua N. H. Plant bZIP protein DNA binding specificity. J Mol Biol. 1993 Apr 20;230(4):1131–1144. doi: 10.1006/jmbi.1993.1230. [DOI] [PubMed] [Google Scholar]
  12. Katagiri F., Yamazaki K., Horikoshi M., Roeder R. G., Chua N. H. A plant DNA-binding protein increases the number of active preinitiation complexes in a human in vitro transcription system. Genes Dev. 1990 Nov;4(11):1899–1909. doi: 10.1101/gad.4.11.1899. [DOI] [PubMed] [Google Scholar]
  13. Lam E., Benfey P. N., Gilmartin P. M., Fang R. X., Chua N. H. Site-specific mutations alter in vitro factor binding and change promoter expression pattern in transgenic plants. Proc Natl Acad Sci U S A. 1989 Oct;86(20):7890–7894. doi: 10.1073/pnas.86.20.7890. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lam E., Chua N. H. ASF-2: a factor that binds to the cauliflower mosaic virus 35S promoter and a conserved GATA motif in Cab promoters. Plant Cell. 1989 Dec;1(12):1147–1156. doi: 10.1105/tpc.1.12.1147. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lam E., Chua N. H. GT-1 binding site confers light responsive expression in transgenic tobacco. Science. 1990 Apr 27;248(4954):471–474. doi: 10.1126/science.2330508. [DOI] [PubMed] [Google Scholar]
  16. Lohmer S., Maddaloni M., Motto M., Di Fonzo N., Hartings H., Salamini F., Thompson R. D. The maize regulatory locus Opaque-2 encodes a DNA-binding protein which activates the transcription of the b-32 gene. EMBO J. 1991 Mar;10(3):617–624. doi: 10.1002/j.1460-2075.1991.tb07989.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Memelink J., Hoge J. H., Schilperoort R. A. Cytokinin stress changes the developmental regulation of several defence-related genes in tobacco. EMBO J. 1987 Dec 1;6(12):3579–3583. doi: 10.1002/j.1460-2075.1987.tb02688.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Salinas J., Oeda K., Chua N. H. Two G-box-related sequences confer different expression patterns in transgenic tobacco. Plant Cell. 1992 Dec;4(12):1485–1493. doi: 10.1105/tpc.4.12.1485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Schindler U., Beckmann H., Cashmore A. R. TGA1 and G-box binding factors: two distinct classes of Arabidopsis leucine zipper proteins compete for the G-box-like element TGACGTGG. Plant Cell. 1992 Oct;4(10):1309–1319. doi: 10.1105/tpc.4.10.1309. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Schmidt R. J., Ketudat M., Aukerman M. J., Hoschek G. Opaque-2 is a transcriptional activator that recognizes a specific target site in 22-kD zein genes. Plant Cell. 1992 Jun;4(6):689–700. doi: 10.1105/tpc.4.6.689. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Singh K., Dennis E. S., Ellis J. G., Llewellyn D. J., Tokuhisa J. G., Wahleithner J. A., Peacock W. J. OCSBF-1, a maize ocs enhancer binding factor: isolation and expression during development. Plant Cell. 1990 Sep;2(9):891–903. doi: 10.1105/tpc.2.9.891. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Skriver K., Mundy J. Gene expression in response to abscisic acid and osmotic stress. Plant Cell. 1990 Jun;2(6):503–512. doi: 10.1105/tpc.2.6.503. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Vancanneyt G., Schmidt R., O'Connor-Sanchez A., Willmitzer L., Rocha-Sosa M. Construction of an intron-containing marker gene: splicing of the intron in transgenic plants and its use in monitoring early events in Agrobacterium-mediated plant transformation. Mol Gen Genet. 1990 Jan;220(2):245–250. doi: 10.1007/BF00260489. [DOI] [PubMed] [Google Scholar]
  24. Vincent J. P., Kassis J. A., O'Farrell P. H. A synthetic homeodomain binding site acts as a cell type specific, promoter specific enhancer in Drosophila embryos. EMBO J. 1990 Aug;9(8):2573–2578. doi: 10.1002/j.1460-2075.1990.tb07438.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Yamazaki K., Katagiri F., Imaseki H., Chua N. H. TGA1a, a tobacco DNA-binding protein, increases the rate of preinitiation complex formation in a plant in vitro transcription system [corrected]. Proc Natl Acad Sci U S A. 1990 Sep;87(18):7035–7039. doi: 10.1073/pnas.87.18.7035. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Plant Cell are provided here courtesy of Oxford University Press

RESOURCES