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Characterization of the expression of a wheat cystatin gene during caryopsis development

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Abstract

A cDNA coding for phytocystatin, a protease inhibitor, was isolated from wheat embryos by differential display RT-PCR and the corresponding full-length cDNA (named WC5 for wheat cystatin gene 5) subsequently obtained by RACE. The deduced primary sequence of the protein suggests the presence of a 28 amino acid N-terminal signal sequence and a 100 amino acid mature protein containing the three consensus motifs known to interact with the active site of cysteine peptidases. Northern and western analysis revealed a spatio-temporal pattern of the cystatin gene expression during caryopse development. In the embryo, WC5 was only expressed during early embryogenesis whereas, in seed covering layers, WC5 expression was restricted to the maturation stage of grain development. In addition, immunolocalization experiments showed that cystatin accumulated in the aleurone layer of the maturating seed and in the parenchymal tissues of the embryo scutellum. A recombinant form of the wheat cystatin was shown to be able to inhibit peptidase activities present in whole seed protein extracts. In addition, immunological techniques allowed us to identify two putative target peptidases. The possible roles of the cystatin protein are discussed in relation with tissular localization and putative peptidase targets during seed maturation.

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References

  • Abe, K., Emori, Y., Kondo, H., Suzuki, K. and Arai, S. 1987. Molecular cloning of a cysteine proteinase inhibitor of rice (oryzacystatin). J. Biol. Chem. 262: 16793–16797.

    Google Scholar 

  • Abe, M., Abe, K., Kuroda, M. and Arai, S. 1992. Corn kernel cysteine proteinase inhibitor as a novel cystatin superfamily member of plant origin. Eur. J. Biochem. 209: 933–937.

    Google Scholar 

  • Abe, Y., Shirane, K., Yokosawa, H., Matsushita, H., Mitta, M., Kato, I. and Ishii, S. 1993. Asparaginyl endopeptidase of jack bean seeds. J. Biol. Chem. 268: 3525–3529.

    Google Scholar 

  • Abe, M., Abe, K., Iwabuchi, K., Domoto, C. and Arai, S. 1994. Corn cystatin I expressed in Escherichia coli: investigation of its inhibitory profile and occurrence in corn kernels. J. Biochem. 116: 488–492.

    Google Scholar 

  • Abe, M., Abe, K., Domoto, C. and Arai, S. 1995. Two distinct species of corn cystatin in corn kernels. Biosci. Biotechnol. Biochem. 59: 756–758.

    Google Scholar 

  • Afonso, S., Romagnano, L. and Babiaz, B. 1997. The expression and function of cystatin C and cathepsin B and cathepsin L during mouse implantation and placentation. Development 124: 3415–3425.

    Google Scholar 

  • Alvarez-Fernandez, M., Barret, A.J., Gerhartz, B., Dando, P.M., Ni, J. and Abrahamson, M. 1999. Inhibition of mammalian legumain by some cystatins is due to a novel second reactive site. J. Biol. Chem. 274: 19195–19203.

    Google Scholar 

  • Beers, E.P., Woffenden, B.J. and Zhao, C. 2000. Plant proteolytic enzymes: possible role during programmed cell death. PlantMol. Biol. 44: 399–415.

    Google Scholar 

  • Berger, D. and Altmann, T. 2000. A subtilisin-like serine protease involved in the regulation of stomatal density and distribution in Arabidopsis thaliana. Genes Dev. 14: 1119–1131.

    Google Scholar 

  • Bode, W., Engh, R., Musil, D., Thiele, U., Huber, R., Karshikov, A., Brzin, J., Kos, J. and Turk, V. 1988. The 2.0 Å X-ray crystal structure of chicken egg white cystatin and its possible mode of interaction with cysteine proteinases. EMBO J. 7: 2593–2599.

    Google Scholar 

  • Bosme, M., Weideman, F. and Olsen, O.-A. 1992. Endosperm differentiation in barley wild-type and sex mutants. Plant J. 2: 661–674.

    Google Scholar 

  • Botella, M.A., Xu, Y., Prabha, T.N., Zhao, Y., Narasimhan, M.L., Wilson, K.A., Nielsen, S.S., Bressan, R.A. and Hasegawa, P.M. 1996. Differential expression of soybean proteinase inhibitor genes during development and in response to wounding and methyl jasmonate. Plant Physiol. 112: 1201–1210.

    Google Scholar 

  • Callis, J. 1995. Regulation of protein degradation. Plant Cell 7: 845–857.

    Google Scholar 

  • Cejudo, F.J., Murphy, G., Chinoy, C. and Baulcombe, D.C. 1992. A gibberellin-regulated gene from wheat with sequence homology to cathepsin B of mammalian cells. Plant J. 2: 937–948.

    Google Scholar 

  • Chen, F. and Foolad, M.R. 1997. Molecular organization of a gene in barley which encodes a protein similar to aspartic protease and its specific expression in nucellar cells during degeneration. Plant Mol Biol. 35: 821–831.

    Google Scholar 

  • Corre, F., Henry, Y., Rode, A. and Hartmann, C. 1996 Em gene expression during somatic embryogenesis in the monocot Triticum aestivum L. Plant Sci. 117: 139–149.

    Google Scholar 

  • Dixon, M. and Webb, E.C. 1979. The Enzymes. Longmans, London, pp. 332–380.

    Google Scholar 

  • Dominguez, F. and Cejudo F.J. 1995. Pattern of endoproteolysis following wheat grain germination. Physiol. Plant. 95: 253–259.

    Google Scholar 

  • Dominguez, F. and Cejudo F.J. 1996. Characterization of the endoproteases appearing during wheat grain development. Plant Physiol. 112: 1211–1217.

    Google Scholar 

  • Dominguez, F. and Cejudo F.J. 1998. Germination-related genes encoding proteolytic enzymes are expressed in the nucellus of developing wheat grains. Plant J. 15: 569–574.

    Google Scholar 

  • Fernandez, K.V.S., Sabelli, P.A., Barratt, D.H.P., Richardson, M., Xavier-Filho, J. and Shewry, P.R. 1993. The resistance of cowpea seeds to bruchid beetles is not related to levels of cysteine proteinase inhibitors. Plant Mol. Biol. 23: 215–219.

    Google Scholar 

  • Joshi, C.P., Zhou, H., Huang, X. and Chiang V.L. 1997. Context sequences of translation initiation codon in plants. Plant Mol. Biol. 35: 993–1001.

    Google Scholar 

  • Hara-Nishimura, I., Inoue, K. and Nishimura, M. 1991. A unique vacuolar processing enzyme responsible for conversion of several pro-protein precursors into the mature form. FEBS Lett. 294: 89–93.

    Google Scholar 

  • Koiwa, H., Bressan, R.A. and Hasegawa, P.M. 1997 Regulation of protease inhibitors and plant defense. Trends Plant Sci. 2: 379–384.

    Google Scholar 

  • Kondo, H., Abe, K., Nishimura, I., Watanabe, H., Emori, Y. and Arai, S. 1990. Two distinct cystatin species in rice seeds with different specificities against cysteine proteinases. J. Biol. Chem. 265: 15832–15837.

    Google Scholar 

  • Kos, J., Werle, B., Lah T. and Brunner N. 2000. Cysteine proteinases and their inhibitors in extracellular fluids: markers for diagnosis and prognosis in cancer. Int. J. Biol. Markers 15: 84–89.

    Google Scholar 

  • Kuroda, M., Kiyosaki, T., Matsumoto, I., Misaka, T., Arai, S. and Abe K. 2001. Molecular cloning, characterization, and expression of wheat cystatins. Biosci. Biotechnol. Biochem. 65: 22–28.

    Google Scholar 

  • Leplé, J.C., Bonadé-Bottino, M., Augustin, S., Pilate, G., Lê Tran, V.D., Deplanque, A., Cornu, D. and Jouanin, L. 1995. Toxicity to Chrysomela tremulae (Coleoptera: Chrysomelidae) of transgenic poplars expressing a cysteine proteinase inhibitor. Mol. Breed. 1: 319–328.

    Google Scholar 

  • Linnestad, C., Doan, D.N.P., Brown, R.C.X., Lemmon, B.E., Meyer, D.J., Jung, R. and Olsen, O.-A. 1998. Nucellain, a barley homolog of the dicot vacuolar-processing proteases, is localized in nucellar cell walls. Plant Physiol. 118: 1169–1180.

    Google Scholar 

  • Machleidt, W., Thiele, U., Laber, B., Assfalg-Machleidt, I., Esterl, A., Wiegand, G., Kos, J., Turk, V. and Bode, W. 1989. Mechanism of inhibition of papain by chicken egg white cystatin. FEBS Lett. 243: 234–238.

    Google Scholar 

  • Margis, R., Reis, E.M. and Villeret, V. 1998. Structural and phylogenetic relationships among plant and animal cystatins. Arch. Biochem. Biophys. 359: 24–30.

    Google Scholar 

  • Michaud, D., Faye, L., and Yelle, S. 1993. Electrophoretic analysis of plant cysteine and serine proteinases using gelatin-containing polyacrylamide gels and class specific proteinase inhibitors. Electrophoresis 14: 94–98.

    Google Scholar 

  • Michaud, D., Nguyen-Quoc B. and Yelle, S. 1994 Production of oryzacystatins I and II in Escherichia coli using the glutathione S-transferase gene fusion system. Biotechnol. Prog. 10: 155–159.

    Google Scholar 

  • Misaka, T., Kuroda, M., Iwabuchi, K., Abe, K. and Arai S. 1996. Soyacystatin, a novel cysteine proteinase inhibitor in soybean, is distinct in protein structure and gene organization from other cystatins of animal and plant origin. Eur. J. Biochem. 240: 609–614.

    Google Scholar 

  • Morrison, I.N., O'Brien, T.P. and Kuo, J. 1978. Initial cellularization and differenciation of the aleurone cells in the ventral region of the developing wheat grain. Planta 140: 19–30.

    Google Scholar 

  • Ojima, A., Shiota, H., Higashi, K., Kamada, H., Shimma, Y.-I., Wada,M. and Satoh, S. 1997. An extracellular insoluble inhibitor of cysteine proteinases in cell cultures and seeds of carrot. Plant Mol. Biol. 34: 99–109.

    Google Scholar 

  • Pernas, M., Sanchez-Monge, R. and Salcedo, G. 2000. Biotic and abiotic stress can induce cystatin expression in chestnut. FEBS Lett. 467: 206–210.

    Google Scholar 

  • Ryan, S., Laing, W.A. and McManus, M.T. 1998. A cysteine proteinase inhibitor purified from apple fruit. Phytochemistry 49: 957–963.

    Google Scholar 

  • Solomon, M., Belenghi, B., Delledonne, M., Menachem, E. and Levine, A. 1999. The involvement of cysteine proteases and protease inhibitor genes in the regulation of programmed cell death in plants. Plant Cell 11: 431–443.

    Google Scholar 

  • Töhönen, V., Österlund, C. and Nordqvist, K. 1998. Testatin: a cystatin-related gene expressed during early testis development. Proc. Natl. Acad. Sci. USA 95: 14208–14213.

    Google Scholar 

  • Turk, V. and Bode, W. 1991. The cystatins: protein inhibitors of cysteine proteinases. FEBS Lett. 285: 213–219.

    Google Scholar 

  • Vierstra, R.D. 1996. Proteolysis in plants: mechanisms and functions. Plant Mol. Biol. 32: 275–302.

    Google Scholar 

  • Womack, J.S., Randall, J. and Kemp, J.D. 2000. Identification of a signal peptide for oryzacystatin-I. Planta 210: 844–847.

    Google Scholar 

  • Young, T.E. and Gallie D.R. 2000. Programmed cell death during endosperm development. Plant Mol. Biol. 44: 283–301.

    Google Scholar 

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Authors and Affiliations

  1. Institut de Biotechnologie des Plantes, Bâtiment 630, Université Paris Sud, 91405, Orsay, cedex, France

    Fabienne Corre-Menguy, Christelle Mazubert, Jean Vidal, Christine Lelandais-Brière, Gisele Torres, André Rode & Caroline Hartmann

  2. Centro de Investigaciones Cientificas Isla de la Cartuja, Instituto de Bioquimica Vegetal y Fotosintesis, avda Americo Vespucio s/n, 41092, Sevilla, Spain

    Francisco J. Cejudo

Authors
  1. Fabienne Corre-Menguy
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  2. Francisco J. Cejudo
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  3. Christelle Mazubert
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  4. Jean Vidal
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  5. Christine Lelandais-Brière
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  7. André Rode
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  8. Caroline Hartmann
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Corre-Menguy, F., Cejudo, F.J., Mazubert, C. et al. Characterization of the expression of a wheat cystatin gene during caryopsis development. Plant Mol Biol 50, 687–698 (2002). https://doi.org/10.1023/A:1019906031305

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