Plant Molecular Biology

, Volume 27, Issue 6, pp 1197–1204

Structure and induction pattern of a novel proteinase inhibitor class II gene of tobacco

  • Teresa Balandin
  • Chris van der Does
  • José-Maria Bellés Albert
  • John F. Bol
  • Huub J. M. Linthorst
Short Communication


A cDNA and a corresponding genomic clone encoding a protein with partial identity to type II proteinase inhibitors from potato, tomato and Nicotiana alata, were isolated from tobacco libraries. The protein of 197 amino acids contains a putative signal peptide of 24 residues and three homologous domains, each with a different reactive site. The tobacco PI-II gene is not expressed in leaves of healthy plants, but is locally induced in leaves subjected to different types of stress (TMV infection, wounding, UV irradiation) and upon ethephon treatment. As opposed to the analogous PI-II genes of potato and tomato, the tobacco gene is not systemically induced by wounding or pathogenic infection. A far-upstream region in the PI-II promoter, containing various direct and indirect repeats, shares considerable sequence similarity to a similar region in the stress-inducible Cu/Zn-superoxide dismutase gene of N. plumbaginifolia.

Key words

pathogenesis-related proteins proteinase inhibitor signal transduction wounding 


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  1. 1.
    Atkinson AH, Heath RL, Simpson RJ, Clarke AE, Anderson MA: Proteinase inhibitors in Nicotiana alata stigmas are derived from a precursor protein which is processed into 5 homologous inhibitors. Plant Cell 5: 203–213 (1993).PubMedGoogle Scholar
  2. 2.
    Bowles DJ: Defense-related proteins in higher plants. Annu Rev Biochem 59: 873–907 (1990).CrossRefPubMedGoogle Scholar
  3. 3.
    Brederode FT, Linthorst HJM, Bol JF: Differential induction of acquired resistance and PR gene expression in tobacco by virus infection, ethephon treatment, UV light and wounding. Plant Mol Biol 17: 1117–1125 (1991).PubMedGoogle Scholar
  4. 4.
    Bryant J, Green TR, Gurusaddaiah T, Ryan CA: Protein-ase inhibitor II from potatoes: isolation and characterization of its protomer components. Biochemistry 15: 3418–3424 (1976).PubMedGoogle Scholar
  5. 5.
    Dougherty WG, Semler BL: Expression of virus-encoded proteinases: functional and structural similarities with cellular enzymes. Microbiol Rev 57: 781–822 (1993).PubMedGoogle Scholar
  6. 6.
    Graham JS, Pearce G, Merryweather J, Titani K, Ericsson LH, Ryan CA: Wound-induced proteinase inhibitors from tomato leaves. II. The cDNA-deduced primary structure of pre-inhibitor II. J Biol Chem 260: 6561–6564 (1985).PubMedGoogle Scholar
  7. 7.
    Greenblatt HM, Ryan CA, James MNG: Structure of the complex of Streptomyces griseus proteinase b and polypeptide chymotrypsin inhibitor-I from Russet Burbank potato tubers at 2.1 Å resolution. J Mol Biol 205: 201–228 (1989).PubMedGoogle Scholar
  8. 8.
    Green TR, Ryan CA: Wound-induced proteinase inhibitors in plant leaves: a possible defense mechanism against insects. Science 175: 776–777 (1972).Google Scholar
  9. 9.
    Hass GM, Hermodson MA, Ryan CA, Gentry L: Primary structures of two low molecular weight proteinase inhibitors from potatoes. Biochemistry 21: 752–756 (1982).PubMedGoogle Scholar
  10. 10.
    Heitz T, Geoffroy P, Stintzi A, Fritig B, Legrand M: cDNA cloning and gene expression analysis of the microbial proteinase inhibitor of tobacco. J Biol Chem 268: 16987–16992 (1993).PubMedGoogle Scholar
  11. 11.
    Herouart DM, VanMontagu MM, Inze DM: Redox-activated expression of the cytosolic copper/zinc superoxide dismutase gene in Nicotiana. Proc Natl Acad Sci USA 90: 3108–3112 (1993).PubMedGoogle Scholar
  12. 12.
    Keil M, Sanchez-Serrano J, Schell J, Willmitzer L: Primary structure of a proteinase inhibitor II gene from potato (Solanum tuberosum). Nucl Acids Res 14: 5641–5650 (1986).PubMedGoogle Scholar
  13. 13.
    Laskowski MJr, Kato I, Ardelt W, Cook J, Denton A, Empie MW, Kohr WJ, Park SJ, Parks K, et al.: Ovomucoid third domains from 100 avian species: isolation, sequences, and hypervariability of enzyme-inhibitor contact residues. Biochemistry 26: 202–221 (1987).PubMedGoogle Scholar
  14. 14.
    Linthorst HJM, vanLoon LC, vanRossum CMA, Mayer A, Bol JF, vanRoekel JSC, Meulenhoff EJS, Cornelissen BJC: Analysis of acidic and basic chitinases from tobacco and petunia and their constitutive expression in transgenic tobacco. Mol Plant-Microbe Interact 3: 252–258 (1990).PubMedGoogle Scholar
  15. 15.
    Linthorst HJM, Brederode FT, van derDoes C, Bol JF: Tobacco proteinase inhibitor-I genes are locally, but not systemically induced by stress. Plant Mol Biol 21: 985–992 (1993).PubMedGoogle Scholar
  16. 16.
    Palm CJ, Costa MA, An G, Ryan CA: Wound-inducible nuclear protein binds DNA fragments that regulate a proteinase inhibitor II gene from potato. Proc Natl Acad Sci USA 87: 603–607 (1990).PubMedGoogle Scholar
  17. 17.
    Pearce G, Johnson S, Ryan CA: Purification and characterization from tobacco (Nicotiana tabacum) leaves of 6 small, wound-inducible, proteinase isoinhibitors of the potato inhibitor-II family. Plant Physiol 102: 639–644 (1993).CrossRefPubMedGoogle Scholar
  18. 18.
    Pena-Cortes H, Sánchez-Serrano J, Rocha-Sosa M, Willmitzer L: Systemic induction of proteinase inhibitor-II gene expression in potato plants by wounding. Planta 174: 84–89 (1988).Google Scholar
  19. 19.
    Plunkett G, Senear DF, Zuroske G, Ryan CA: Protein-ase inhibitor I and II from leaves of wounded tomato plants: purification and properties. Arch Biochem Biophys 213: 463–472 (1982).PubMedGoogle Scholar
  20. 20.
    Richardson M: The complete amino acid sequence and the trypsin reactive (inhibitory) site of the major protein-ase inhibitor from the fruits of aubergine (Solanum melongena L.). FEBS Lett 104: 322–326 (1979).CrossRefPubMedGoogle Scholar
  21. 21.
    Ryan CA: Protease inhibitors in plants: genes for improving defenses against insects and pathogens. Annu Rev Phytopath 28: 425–449 (1990).CrossRefGoogle Scholar
  22. 22.
    Sánchez-Serrano J, Schmidt R, Schell J, Willmitzer L: Nucleotide sequence of proteinase inhibitor II encoding cDNA of potato (Solanum tuberosum) and its mode of expression. Mol Gen Genet 203: 15–20 (1986).CrossRefGoogle Scholar
  23. 23.
    Sánchez-Serrano JJ, Keil M, O'Connor A, Schell J, Willmitzer L: Wound-induced expression of a potato proteinase inhibitor II gene in transgenic tobacco plants. EMBO J 6: 303–306 (1987).Google Scholar
  24. 24.
    Taylor BH, Young RJ, Scheuring CF: Induction of a proteinase inhibitor II-class gene by auxin in tomato roots. Plant Mol Biol 23: 1005–1014 (1993).PubMedGoogle Scholar
  25. 25.
    Thornburg RW, An G, Cleveland TE, Johnson R, Ryan CA: Wound-inducible expression of a potato inhibitor II-chloramphenicol acetyltransferase gene fusion in transgenic tobacco plants. Proc Natl Acad Sci USA 84: 744–748 (1987).Google Scholar
  26. 26.
    Wildon DC, Thain JF, Minchin PEH, Gubb IR, Reilly AJ, Skipper YD, Doherty HM, O'Donnell PJ, Bowles DJ: Electrical signalling and systemic proteinase inhibitor induction in the wounded plant. Nature 360: 62–65 (1992).CrossRefGoogle Scholar

Copyright information

© Kluwer Academic Publishers 1995

Authors and Affiliations

  • Teresa Balandin
    • 1
  • Chris van der Does
    • 1
  • José-Maria Bellés Albert
    • 1
  • John F. Bol
    • 1
  • Huub J. M. Linthorst
    • 1
  1. 1.Institute of Molecular Plant SciencesGorlaeus LaboratoriesLeidenThe Netherlands
  2. 2.Department of Molecular MicrobiologyGroningen UniversityHarenThe Netherlands
  3. 3.Dep. Biotecnología, E.T.S.I. AgrónomosUniv. PolitécnicaValenciaSpain

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