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Molecular identification and expression of the peroxidase responsible for the oxidative burst in French bean (Phaseolus vulgaris L.) and related members of the gene family

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Abstract

Molecular characterization has been accomplished for five members of the peroxidase gene family in French bean. The most important of these, designated FBP1, corresponds to the isoform believed to be responsible for the apoplastic oxidative burst demonstrated by suspension-cultured cells in response to fungal elicitor. Identification was made by a complete match of six peptide sequences derived from the native protein to the translated sequence of the cDNA. Modelling of the surface structure in comparison with two other members of the peroxidase family did not reveal any unusual features which might account for its role in the oxidative burst. However, FBP1 when expressed in Pichia pastoris generated H2O2 using cysteine at pH 7.2, a specific property of the native protein when isolated from suspension-cultured cells. FBP1, together with other members of the family, were all induced in cell cultures by elicitor action although they all showed some expression in non-induced cultured cells. They were also expressed in all tissues examined with varying levels of intensity of detection in northern blots. This was confirmed by in situ hybridization and FBP1 expression was confirmed in tissues where it has been previously detected by immunolocalization methods. Assigning roles to individual peroxidases is an important goal and molecular identification of the oxidative burst peroxidase allows further exploration of the relative roles of the different systems involved in generating reactive oxygen species.

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References

  • Altschul, S.F., Madden, T.L., Schaffer, A.A., Zhang J., Zhang, Z., Miller, W. and Lipman, D.J. 1997. Capped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucl. Acids Res. 25: 3389–3402.

    Google Scholar 

  • Angerer, L.M. and Angerer, R.C. 1991. Localization of mRNAs by in situ hybridization. Meth. Cell Biol. 35: 37–71.

    Google Scholar 

  • Ausubel, F.M., Brent, R., Kingston, R.E., Moore, D.D., Seidman, J.G., Smith, J.A. and Struhl, K. (Eds.). 1999. Current Protocols in Molecular Biology. John Wiley, New York.

    Google Scholar 

  • Barroso, J.B., Corpas, F.J., Carreras, A., Sandalio, L.M., Valderrama, R., Palma, J.M., Lupianez, J.A. and del Rio, L.A. 1999. Localization of nitric-oxide synthase in plant peroxisomes. J. Biol. Chem. 274: 36729–36733.

    Google Scholar 

  • Bednarek, S.Y. and Raikhel, N.V. 1991. Intracellular trafficking of secretory proteins. Plant Mol. Biol. 20: 133–150.

    Google Scholar 

  • Bednarek, S.Y. and Raikhel, N.V. 1992. The barley lectin carboxyl-terminal propeptide is a vacuolar sorting determinant in plants. Plant Cell 3: 1195–1206.

    Google Scholar 

  • Bernards, M.A., Fleming, W.D., Llewellyn, D.B., Priefer, R., Yang, X., Sabatino, A. and Plourde, G.L. 1999. Biochemical characterisation of the suberisation-associated anionic peroxidase of potato. Plant Physiol. 121: 135–146.

    Google Scholar 

  • Bjellqvist, B., Hughes, G.J., Pasquali, C., Paquet, N., Ravier, F., Sanchez, J.-C., Frutiger, S. and Hochstrasser, D.F. 1993. The focusing positions of polypeptides in immobilized pH gradients can be predicted from their amino acid sequences. Electrophoresis 14: 1023–1031.

    Google Scholar 

  • Blee, K.A. and Anderson, A.J. 1996. Defense-related transcript accumulation in Phaseolus vulgaris L. colonized by the arbuscular mycorrhizal fungus Glomus intraradices Schenck and Smith. Plant Physiol 110: 675–688.

    Google Scholar 

  • Bolwell, G.P. 1996. The origin of the oxidative burst in plants. Biochem. Soc. Trans. 24: 438–442.

    Google Scholar 

  • Bolwell, G.P. 1999. Role of active oxygen species and NO in plant defence responses. Curr. Opin. Plant Biol. 2: 287–294.

    Google Scholar 

  • Bolwell, G.P. and Wojtaszek, P. 1997. Mechanisms for the generation of reactive oxygen species in plant defence ¶ a broad perspective. Physiol. Mol. Plant Path. 51: 347–366.

    Google Scholar 

  • Bolwell, G.P., Butt, V.S., Davies, D.R. and Zimmerlin, A. 1995. The origin of the oxidative burst in plants. Free Radical Res. 23: 517–532.

    Google Scholar 

  • Bolwell, G.P., Davies, D.R., Gerrish, C., Auh, C.-K. and Murphy, T.M. 1998. Comparative biochemistry of the oxidative burst produced by rose and french bean cells reveals two distinct mechanisms. Plant Physiol. 116: 1379–1385.

    Google Scholar 

  • Bolwell, G.P., Blee, K.R., Butt, V.S., Davies, D.R., Gardner, S.L., Gerrish, C., Minibayeva, F., Rowntree, E.G. and Wojtaszek, P. 1999. Recent advances in understanding the origin of the oxidative burst in plant cells. Free Radical Res. 31: S137–S145.

    Google Scholar 

  • Brown, I., Trethowan, J., Kerry, M., Mansfield, J.W. and Bolwell, G.P. 1998. Location of components of the oxidative cross-linking of glycoproteins and callose synthesis in papillae formed during the interaction between non-pathogenic strains of Xanthomonas campestris and french bean mesophyll cells. Plant J. 15: 333–343

    Google Scholar 

  • Cox, K.H., DeLeon, D.V., Angerer, L.M. and Angerer, R.C. 1984. Detection of mRNAs in sea urchin embryos by in situ hybridization using asymmetric RNA probes. Dev. Biol. 101: 485–502.

    Google Scholar 

  • Dixon, R.A. and Lamb, C.J. 1979. Stimulation of de novo synthesis of L-phenylalanine ammonia-lyase in relation to phytoalexin accumulation of Colletotrichum lindemuthianum elicitor-treated cell suspension cultures of French bean (Phaseolus vulgaris). Biochim. Biophys. Acta 586: 453–463.

    Google Scholar 

  • Elliott, K., Butler, W., Dickinson, C.D., Konno, Y., Vedvick, T.S., Fitzmaurice, L. and Mirkov, T.E. 1993. Isolation and characterization of fruit vacuolar invertase genes from two tomato species and temporal differences in mRNA levels during fruit ripening. Plant Mol. Biol. 21: 515–524.

    Google Scholar 

  • el-Turk, J., Asemota, O., Leymarie, J., Sallaud, C., Mesnage, S., Breda, C., Buffard, D., Kondorosi, A. and Esnault, R. 1996. Nucleotide sequences of four pathogen-induced alfalfa peroxidase-encoding cDNAs. Gene 170: 213–216.

    Google Scholar 

  • Fujiyama, K., Takemura, H., Shibayama, S., Kobayashi, K., Choi, J.K., Shinmyo, A., Takano, M., Yamada, Y. and Okada, H. 1988. Structure of the horseradish peroxidase isozyme C genes. Eur. J. Biochem. 3: 681–687.

    Google Scholar 

  • Gazarian, I.G., Lagrimini, L.M., Mellon, F.A., Naldrett, M.J., Ashby, G.A., Thorneley, R.N. 1998. Identification of skatolyl hydroperoxide and its role in the peroxidase-catalysed oxidation of indol-3-yl acetic acid. Biochem. J. 333: 223–232

    Google Scholar 

  • Giannino, D.S., Manfiotti, G. and Schneider, C. 1988. A one-tube plasmid DNA mini-preparation suitable for sequencing. Nucl. Acids Res. 16: 9878.

    Google Scholar 

  • Hiraga, S., Ito, H., Matsui, H., Honma, M. and Ohashi, Y. 1999. cDNA sequences for two novel tobacco peroxidase isoenzymes. Plant Physiol. 120: 1205.

    Google Scholar 

  • Huang, W.M., Gibson, S.J., Facer, P., Gu, J. and Polak, J.M. 1983. Improved section adhesion for Immunocytochemistry using high molecular weight polymers of L-lysine as a slide coating. Histochemistry 77: 275–279.

    Google Scholar 

  • Keller, T., Damude, H.G., Werner, D., Doerner P., Dixon, R.A. and Lamb, C.J. 1998. A plant homolog of the neutrophil NADPH-oxidase gp91 phox subunit gene encodes a plasma membrane protein with Ca 2+ binding motifs. Plant Cell 10: 255–266

    Google Scholar 

  • Klann, E., Yelle, S. and Bennett, A.B., 1992. Tomato fruit acid invertase complementary DNA. Plant Physiol. 99: 351–353.

    Google Scholar 

  • Lamb, C.J. and Dixon, R.A. 1997. The oxidative burst in plant disease resistance. Annu. Rev. Plant Physiol. Mol. Biol. 48: 251–275.

    Google Scholar 

  • Lewis, N.G., Davin, L.B. and Sarkanen, S. 1999. The nature and function of lignins. In: P.M. Pinto (Ed.) Comprehensive Natural Product Chemistry, Vol. 3, Elsevier, Amsterdam, pp. 617–745.

    Google Scholar 

  • Logemann, J., Schell, J. and Willmitzer, L. 1987. Improved method for the isolation of RNA from plant tissues. Anal. Biochem. 163: 16–20.

    Google Scholar 

  • Lopez-Huertas, E., Corpas, F.J., Sandalio, L.M. and del Rio, L.A. 1999. Characterization of membrane polypeptides from pea leaf peroxisomes involved in superoxide radical generation. Biochem. J. 337: 531–536.

    Google Scholar 

  • Luthe, D.S. and Quatrano, R.S. 1980. Transcription in isolated wheat nuclei. I. Isolation of nuclei and elimination of endogenous ribonuclease activity. Plant Physiol. 65: 305–308.

    Google Scholar 

  • Matsuoka, K. and Nakamura, K. 1991. Propeptide of a precursor to a plant vacuolar protein required for vacuolar targeting. Proc. Natl. Acad. Sci. USA 88: 834–838.

    Google Scholar 

  • Neuhaus, J.-M., Sticher, L., Meins, F. and Boller, T. 1991. A short C-terminal sequence necessary and sufficient for the targeting of chitinase to the plant vacuole. Proc. Natl. Acad. Sci. USA 88: 10362–10366.

    Google Scholar 

  • Ostergaard, L., Abelskov, A.K., Mattsson, O. and Welinder, K.G. 1996. Structure and organ specificity of an anionic peroxidase from Arabidopsis thaliana cell suspension culture. FEBS Lett. 398: 243–247.

    Google Scholar 

  • Ostergaard, L., Pedersen, A.G., Jespersen, H.M., Brunak, S., Welinder, K.G. 1998. Computational analyses and annotations of the Arabidopsis peroxidase gene family. FEBS Lett. 433: 98–102.

    Google Scholar 

  • Ostergaard, L., Teilum, K., Mirza, O., Petersen, M., Welinder, K.G., Mundy, J., Gajhede, M. and Hendriksen, A. 2000. Arabidopsis ATPA2 peroxidase: expression and high-resolution structure of a plant peroxidase with implications for lignification. Plant Mol. Biol. 44: 231–243.

    Google Scholar 

  • Rodgers, M.W., Zimmerlin, A., Werck-Reichhart, D. and Bolwell, G.P. 1993 Microsomally associated heme proteins from French bean: characterization of the cytochrome P450 cinnamate 4-hydroxylase and two peroxidases. Arch. Biochem. Biophys. 304: 74–80.

    Google Scholar 

  • Sambrook, J., Fritsch, E.F. and Maniatis, T. 1989. Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory Press, Plainview, NY.

    Google Scholar 

  • Smith, C.G., Rodgers, M.W., Zimmerlin, A., Ferdinando, D. and Bolwell, G.P. 1994. Tissue and subcellular immunolocalisation of enzymes of lignin synthesis in differentiating and wounded hypocotyl tissue of French bean (Phaseolus vulgaris L.). Planta 192: 155–164.

    Google Scholar 

  • Torre, M.A., Onuchi, H., Hamada, S., Machida, C., Hammond-Kosak, K.E. and Jones, J.D.G. 1998. Six Arabidopsis thaliana homologues of the human respiratory burst oxidase (gp 91¶phox). Plant J. 14: 365–370.

    Google Scholar 

  • von Heijne, G. 1983. Patterns of amino acids near signal sequence cleavage sites. Eur. J. Biochem. 133: 17–21.

    Google Scholar 

  • von Heijne, G. 1986. A new method for predicting signal sequence cleavage sites. Nucl. Acids Res. 14: 4683–4690.

    Google Scholar 

  • Welinder, K.G. 1991. The plant peroxidase superfamily. In: J. Lo-barzewski, H. Greppin, C. Penel and T. Gaspar (Eds.) Biochemical, Molecular, and Physiological Aspects of Plant Peroxidases, University of Geneva Press, Geneva, Switzerland, pp. 3–13.

    Google Scholar 

  • Winther, J.R. and Sorensen, P. 1991. Propeptide of carboxy-peptidase Y provides a chaperone-like function as well as inhibition of the enzymatic activity. Proc. Natl. Acad. Sci. USA 88: 9330–9334.

    Google Scholar 

  • Wojtaszek, P., Trethowan, J. and Bolwell, G.P. 1997. Reconstitution in vitro of the components and conditions required for the oxidative cross-linking of extracellular proteins in French bean (Phaseolus vulgaris L.) FEBS Lett. 405: 95–98.

    Google Scholar 

  • Zimmerlin, A., Wojtaszek, P. and Bolwell, G.P. 1994. Synthesis of dehydrogenation polymers of ferulic acid with high specificity by a purified cell-wall peroxidase from French bean (Phaseolus vulgaris L.). Biochem. J. 299: 747–753.

    Google Scholar 

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Blee, K.A., Jupe, S.C., Richard, G. et al. Molecular identification and expression of the peroxidase responsible for the oxidative burst in French bean (Phaseolus vulgaris L.) and related members of the gene family. Plant Mol Biol 47, 607–620 (2001). https://doi.org/10.1023/A:1012307324782

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