Abstract
Elicitins form a family of 10-kDa holoproteins secreted by various Phytophthora species. The large-scale purification of parasiticein, a novel elicitin secreted by P. parasitica, led to the determination of its sequence. We have compared the necrotic activities and the primary and secondary structures (determined through circular dichroism) of four elicitins. On tobacco plants, they could be classified into two classes: a, comprising capsicein and parasiticein (less necrotic), and β, comprising cryptogein and cinnamomin (very toxic with a necrosis threshold of 0.1 μg per leaf). The features of elicitin structure which might be involved in the interaction of elicitins with the leaf target cells and that could explain the different necrosis-inducing properties of the two proteins are investigated. About 75% sequence identity was observed between the four elicitins: only two short terminal regions are heterologous, while the central core is mainly conserved. The circular-dichroism spectra showed that the secondary structure of the elicitins was largely conserved. All of them consisted of approx. 50% α-helix with little or no β-structure. Comparisons of the complete sequences, amino-acid compositions, isoelectric points, hydropathy indices and the secondary-structure predictions correlated with the necrotic classification. Alpha elicitins corresponded to acidic molecules with a valine residue at position 13, while β elicitins were basic with a lysine at this position, which appeared to be a putative active site responsible for necrosis induction.
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Abbreviations
- CD:
-
circular dichroism
- RPLC:
-
reversed-phase liquid chromatography
- SDS-PAGE:
-
sodium dodecyl sulfate-polyacrylamide gel electrophoresis
References
Baudet, J., Huet, J.C., Jolivet, E., Lesaint, C., Mossé, J., Pernollet, J.C. (1986) Changes in accumulation of seed nitrogen compounds in maize under conditions of sulphur deficiency. Physiol. Plant. 68, 608–614
Beissmann, B., Reisener, H.J. (1990) Isolation and purity determination of a glycoprotein elicitor from wheat stem rust by medium-pressure liquid chromatography. J. Chromatogr. 521, 187–197
Billard, V., Bruneteau, M., Bonnet, P., Ricci, P., Pernollet, J.C., Huet, J.C., Vergne, A., Richard, G., Michel, G. (1988) Chromatographic purification and characterization of elicitors of necrosis on tobacco produced by incompatible Phytophthora species. J. Chromatogr. 44, 87–94
Bonnet, P., Poupet, A., Bruneteau, M. (1985) Toxicité vis-à-vis du tabac des fractions purifiées d'un filtrat de culture de Phytophthora cryptogea Pethyb. & Laff. Agronomie 5, 275–282
Bowles, D.J. (1990) Defense-related proteins in higher plants. Annu. Rev. Biochem. 59, 873–907
Dayhoff, M.O., Schwartz, R.M., Orcutt, B.C. (1978) A model of evolutionary change in proteins. In: Atlas of protein sequence and structure, vol. 5, Suppl. 3, pp. 345–352, National Biomedical Research Foundation, Washington D.C.
Dessen, P., Fondrat, C., Valencien, C., Mugnier, C. (1990) BISANCE: A french service for access to biomolecular sequence databases. CABIOS 6, 355–356
Farmer, E.E., Helgeson, J.P. (1987) An extracellular protein from Phytophthora parasitica var. nicotianæ is associated with stress metabolite accumulation in tobacco callus. Plant Physiol. 85, 733–740
Gamier, J., Osguthorpe, D.J., Robson, B. (1978) Analysis of the accuracy and implications of simple methods for predicting the secondary structure of globular proteins. J. Mol. Biol. 120, 97–120
Henschen, A. (1986) Analysis of cyst(e)ine residues, disulfide bridges, and sulfhydryl groups in proteins. In: Advanced methods in protein microsequence analysis, pp. 244–255, Wittmann-Liebold, B., Salnikow, J., Erdmann, V.A., eds. Springer, Berlin
Hou, K.C., Mandaro, R.M. (1986) Bioseparation by ion exchange cartridge chromatography. Biotechniques 4, 358–367
Huet, J.C., Pernollet, J.C. (1989) Amino acid sequence of cinnamomin, a new member of the elicitin family, and its comparison to cryptogein and capsicein. FEBS Lett. 257, 302–306
Huet, J.C., Sallantin, M., Pernollet, J.C. (1990) Large scale purification of elicitins, protein elicitors secreted by the plant pathogenic fungi Phytophthora. In: Technologies de purification des protéines, vol. 4, pp. 29–34, Briand, Y., Doinel, C., Faure, A., eds. G.R.B.P., Villebon/Yvette, France
Kanehisa, M. (1984) Use of statistical criteria for screening potential homologies in nucleic acid sequences. Nucleic Acids Res. 12, 203–213
Kyte, J., Doolittle, R.F. (1982) A simple method for displaying the hydropathic character of a protein. J. Mol. Biol. 157, 105–132
Parker, J.E., Hahlbrock, K., Scheel, D. (1988) Different cell-wall components from Phytophthora megasperma f. sp. glycinea elicit phytoalexin production in soybean and parsley. Planta 176, 75–82
Plich, M., Rudnicki, R.M. (1979) Studies of the toxins of Phytophthora cactorum pathogenic to apple trees. I. Isolation, some of the properties and activities of a toxin produced by the fungus cultured in vitro. Phytopathology Z. 94, 270–278
Ricci, P., Bonnet, P., Huet, J.C., Sallantin, M., Beauvais-Cante, F., Bruneteau, M., Billard, V., Michel, G., Pernollet, J.C. (1989) Structure and activity of proteins from pathogenic fungi Phytophthora eliciting necrosis and acquired resistance in tobacco. Eur. J. Biochem. 183, 555–563
Sallantin, M., Huet, J.C., Demarteau, C., Pernollet, J.C. (1990) Reassessment of commercially available molecular weight standards for peptide sodium dodecyl sulfate-polyacrylamide gel electrophoresis using electroblotting and microsequencing. Electrophoresis 11, 34–36
Schottens-Toma, I.M.J., De Wit, P.J.G.M. (1988) Purification and primary structure of a necrosis-inducing peptide from the apoplastic fluids of tomato infected with Cladosporium fulvium (syn. Fulvia fulva). Physiol. Mol. Plant Pathol. 33, 59–67
Yang, J.T., Wu, C.-S., Martinez, H.G. (1986) Calculation of protein conformation from circular dichroism. Methods Enzymol. 130, 208–269
Yuen, S.W., Chui, A.H., Wilson, K.J., Yuan, P.M. (1989) Microanalysis of SDS-PAGE electroblotted proteins. Biotechniques 7, 74–83
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The authors are indebted to Dr. A. Van Dorrselaer (Laboratoire de Chimie Organique des Substances Naturelles, Strasbourg, France) for mass-spectrometry measurements. They are grateful to their staff in Versailles, more particularly to Marc Sallantin for electrophoreses, to Françoise Beauvais for biological-activity determinations and to Monique Mansion and Christian Ouali for their skilful technical assistance.
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Nespoulous, C., Huet, JC. & Pernollet, JC. Structure-function relationships of α and β elicitins, signal proteins involved in the plant-Phytophthora interaction. Planta 186, 551–557 (1992). https://doi.org/10.1007/BF00198035
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DOI: https://doi.org/10.1007/BF00198035