Advertisement

Plant Molecular Biology

, Volume 23, Issue 6, pp 1139–1150 | Cite as

Ubiquitous nuclear proteins bind to 5′ upstream region of major Kunitz chymotrypsin inhibitor gene in winged bean

  • Yoshiki Habu
  • Yoichi Sakata
  • Kazuhiro Fukasawa
  • Takeshi Ohno
Research Article

Abstract

Winged bean Kunitz chymotrypsin inhibitor (WCI) accumulates abundantly in seeds and tuberous roots of winged bean plant. In seeds, the WCI mRNA is observed transiently during seed maturation period. The WCI is encoded by a multigene family and the major WCI (WCI-3) is encoded by two nearly identical genes (WCI-3a and WCI-3b genes), in which nucleotide sequences in the 1.1 kb 5′ flanking regions are about 99% homologous to each other and the transcribed regions are completely identical. Here we report the detection of two types of nuclear proteins which bind to the multiple sites in the 5′ upstream region of the WCI-3a gene. One of the proteins, band 1-forming protein, also bound to cauliflower mosaic virus 35S (CaMV35S) promoter, but another protein, band 3-forming protein, did not. DNaseI footprinting analysis showed that these proteins bound to AT-rich upstream regions in the WCI-3a gene. Addition of poly(dA-dT)-poly(dA-dT) to the binding reaction inhibited the formation of the retarded bands, while poly(dI-dC)-poly(dI-dC) did not. In various organs and throughout seed maturation period, proteins with invariable binding specificities were detected, and these binding proteins met some operational criteria for high-mobility-group (HMG) proteins. These results suggest that leguminous seed AT-binding proteins reported on several seed storage protein genes may be HMG-like proteins which are present ubiquitously in plant organs.

Key words

DNA-binding proteins high mobility group proteins HMG proteins Kunitz proteinase inhibitor seed storage protein 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Allen, RD, Bernier, F, Lessard, PA, Beachy, RN: Nuclear factors interact with a soybean β-conglycinin enhancer. Plant Cell 1: 623–631 (1989).Google Scholar
  2. 2.
    Bäumlein, H, Wobus, U, Pustell, J, Kafatos, FC: The legumin gene family: structure of a B type gene of Vicia faba and a possible legumin gene specific regulatory element. Nucl Acids Res 14: 2707–2720 (1986).Google Scholar
  3. 3.
    Benfey, PN, Chua, N-H: The cauliflower mosaic virus 35S promoter: combinatorial regulation of transcription in plants. Science 250: 959–966 (1990).Google Scholar
  4. 4.
    Bradford, MM: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248–254 (1976).Google Scholar
  5. 5.
    Bustin, M, Lehn, DA, Landsman, D: Structural features of the HMG chromosomal proteins and their genes. Biochim Biophys Acta 1049: 231–243 (1990).Google Scholar
  6. 6.
    Bustos, MM, Guiltinan, MJ, Jordano, J, Begum, D, Kalkan, FA, Hall, TC: Regulation of β-glucuronidase expression in transgenic tobacco plants by an A/T-rich, cis-acting sequence found upstream of a French bean β-phaseolin gene. Plant Cell 1: 839–853 (1989).Google Scholar
  7. 7.
    Chen, Z-L, Pan, N-S, Beachy, RN: A DNA sequence element that confers seed-specific enhancement to a constitutive promoter. EMBO J 7: 297–302 (1988).Google Scholar
  8. 8.
    Churchill, MEA, Travers, AA: Protein motifs that recognize structural features of DNA. Trens Biochem Sci 16: 92–97 (1991).Google Scholar
  9. 9.
    Cox, KH, Goldberg, RB: Analysis of plant gene expression. In: Shaw, CH (ed), Plant Molecular Biology: A Practical Approach, pp. 1–35. IRL Press, Oxford (1988).Google Scholar
  10. 10.
    Czarnecka, E, Ingersoll, JC, Gurley, WB: AT-rich promoter elements of soybean heat shock gene Gmhsp 17.5E bind two distinct sets of nuclear proteins in vitro. Plant Mol Biol 19: 985–1000 (1992).Google Scholar
  11. 11.
    Dickinson, CD, Evans, RP, Nielsen, NC: RY repeats are conserved in the 5′-flanking regions of legume seedprotein genes. Nucl Acids Res 16: 371 (1988).Google Scholar
  12. 12.
    Forde, BG, Freeman, J, Oliver, JE, Pineda, M: Nuclear factors interact with conserved A/T-rich elements upstream of a nodule-enhanced glutamine synthetase gene from French bean. Plant Cell 2: 925–939 (1990).Google Scholar
  13. 13.
    Gatehouse, JA, Evans, IM, Croy, RRD, Boulter, D: Differential expression of genes during legume seed development. Phil Trans Roy Soc Lond B314: 367–384 (1986).Google Scholar
  14. 14.
    Goldberg, RB: Regulation of plant gene expression. Phil Trans Roy Soc Lond B314: 343–353 (1986).Google Scholar
  15. 15.
    Goldberg, RB, Barker, SJ, Perez-Grau, L: Regulation of gene expression during plant embryogenesis. Cell 56: 149–160 (1989).Google Scholar
  16. 16.
    Habu, Y, Peyachoknagul, S, Umemoto, K, Sakata, Y, Ohno, T: Structure and regulated expression of Kunitz chymotrypsin inhibitor genes in winged bean (Psophocarpus tetragonolobus (L.) DC.). J Biochem 111: 249–258 (1992).Google Scholar
  17. 17.
    Harada, JJ, Barker, SJ, Goldberg, RB: Soybean β-conglycinin genes are clustered in several DNA regions and are regulated by transcriptional and postranscriptional processes. Plant Cell 1: 415–425 (1989).Google Scholar
  18. 18.
    Jacobsen, K, Laursen, NB, Jensen, EO, Marcker, A, Poulsen, C, Marcker, KA: HMGI-like proteins from leaf and nodule nuclei interact with different AT motifs in soybean nodulin promoters. Plant Cell 2: 85–94 (1990).Google Scholar
  19. 19.
    Jofuku, KD, Goldberg, RB: Analysis of plant gene structure. In: Shaw, CH (ed), Plant Molecular Biology: A Practical Approach, pp. 37–66. IRL Press, Oxford (1988).Google Scholar
  20. 20.
    Jofuku, KD, Okamuro, JK, Goldberg, RB: Interaction of an embryo DNA binding protein with a soybean lectin gene upstream region. Nature 328: 734–737 (1987).Google Scholar
  21. 21.
    Katagiri, F, Chua, N-H: Plant transcription factors: present knowledge and future challenges. Trends Genet 8: 22–27 (1992).Google Scholar
  22. 22.
    Kolodrubetz, D: Consensus sequence for HMG-like DNA binding domains. Nucl Acids Res 18: 5565 (1990).Google Scholar
  23. 23.
    Kruger, W, Herskowitz, I: A negative regulator of HO transcription, SIN1 (SPT2), is a nonspecific DNA-binding protein related to HMG1. Mol Cell Biol 11: 4135–4146 (1991).Google Scholar
  24. 24.
    Laux, T, Goldberg, RB: A plant DNA binding protein shares highly conserved sequence motif with HMG-box proteins. Nucl Acids Res 19: 4769 (1991).Google Scholar
  25. 25.
    Laux, T, Seurinck, J, Goldberg, RB: A soybean embryo cDNA encodes a DNA binding protein with histone and HMG-protein-like domains. Nucl Acids Res 19: 4768 (1991).Google Scholar
  26. 26.
    Lessard, PA, Allen, RD, Bernier, F, Crispino, JD, Fujiwara, T, Beachy, RN: Multiple nuclear factors interact with upstream sequence of differentially regulated β-conglycinin genes. Plant Mol Biol 16: 397–413 (1991).Google Scholar
  27. 27.
    Mitchell, PJ, Tjian, R: Transcriptional regulation in mammalian cells by sequence-specific DNA binding proteins. Science 245: 371–378 (1989).Google Scholar
  28. 28.
    Nielsen, NC, Dickinson, CD, Cho, T-J, Thanh, VH, Scallon, BJ, Ficher, RL, Sims, TL, Drews, GN, Goldberg, RB: Characterization of the glycinin gene family in soybean. Plant Cell 1: 313–328 (1989).Google Scholar
  29. 29.
    Pedersen, TJ, Arwood, LJ, Spiker, S, Guiltinan, MJ, Thompson, WF: High mobility group chromosomal proteins bind to AT-rich tracts flanking plant genes. Plant Mol Biol 16: 95–104 (1991).Google Scholar
  30. 30.
    Peterson, CL, Kruger, W, Herskowitz, I: A functional interaction between the C-terminal domain of RNA polymeraseII and the negative regulator SIN1. Cell 64: 1135–1143 (1991).Google Scholar
  31. 31.
    Peyachoknagul, S, Matsui, T, Shibata, H, Hara, S, Ikenaka, T, Okada, Y, Ohno, T: Sequence and expression of the mRNA encoding the chymotrypsin inhibitor in winged bean (Psophocarpus tetragonolobus (L.) DC.). Plant Mol Biol 12: 51–58 (1989).Google Scholar
  32. 32.
    Ryan, CA: Proteinase inhibitors. In: Marcus, A (ed) The Biochemistry of Plants, Vol. 6, pp. 351–370. Academic Press, New York (1981).Google Scholar
  33. 33.
    Sambrook, J, Fritsch, EF, Maniatis, T: Molecular Cloning: A Laboratory Manual, 2nd ed., pp. E37-E38. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1989).Google Scholar
  34. 34.
    Solomon, MJ, Straus, F, Varshavsky, A: A mammalian high mobility group protein recognizes any stretch of six A-T base pairs in duplex DNA. Proc Natl Acad Sci USA 83: 1276–1280 (1986).Google Scholar
  35. 35.
    Umemoto, K, Habu, Y, Ohno, T: Analysis of differential accumulation of winged bean Kunitz chymotrypsin inhibitor mRNA species by a sequence-specific termination method. Plant Mol Biol 20: 1175–1178 (1992).Google Scholar
  36. 36.
    Walling, L, Drews, GN, Goldberg, RB: Transcriptional and post-transcriptional regulation of soybean seed protein mRNA levels. Proc Natl Acad Sci USA 83: 2123–2127 (1986).Google Scholar

Copyright information

© Kluwer Academic Publishers 1993

Authors and Affiliations

  • Yoshiki Habu
    • 1
  • Yoichi Sakata
    • 1
  • Kazuhiro Fukasawa
    • 1
  • Takeshi Ohno
    • 1
  1. 1.Department of Applied Bioscience, Faculty of AgricultureHokkaido UniversitySapporoJapan

Personalised recommendations