Abstract
The importance of proteolytic enzymes in plant-pest and plant-pathogen interactions has recently been recognized, and control strategies based on their inhibition with protease inhibitors (PIs) have been developed or proposed to control herbivory insects (1), plant parasitic fungi (2,3), and nematodes (4). The various roles of proteases in these organisms and the biochemical pathways affected by their interactions with PIs may differ, but their importance for normal growth and development is now evident. The repressive effects of PIs on insect growth and fecundity, notably, have been documented for several species (1), and evidence for the implication of microbial proteases as phytopathogenic determinants has been reported in several cases (5–13). Based on this information, transformation of plant genomes with PI cDNA clones appears to be an attractive approach for the control of plant pests and pathogens, and several economically important plants expressing exogenous plant PIs have been engineered during the past few years (Table 1). While allowing control of plant pests and pathogens, PIs expressed in transgenic plants may also serve as a source of active inhibitors for the study and the eventual control of some protease-related pathogenic processes in humans. Proteases are important not only in the intracellular regulation of peptides and proteins, but also in the development of several diseases, including tumor metastasis (21), rheumatoid arthritis (22,23), Alzheimer’s disease (24), emphysema (25), pancreatitis (26), and AIDS (27,28).
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References
Hilder, V. A., Gatehouse, A. M. R., and Boulter, D. (1993) Proteinase inhibitor approach, in Transgenic Plants∶ Engineering and Utilization, vol. 1 (Kung, S.-D. and Wu, R., eds.), Academic, New York, pp. 317–338.
Lorito, M., Broadway, R. M., Hayes, C. K., Woo, S. L., Noviello, C., Williams, D. L., and Harman, G. E. (1994) Proteinase inhibitors from plants as a novel class of fungicides. Mol. Plant-Microbe Interact. 7, 525–527.
Dunaevskii, Y. E., Pavlyukova, E. B., Belyakova, G. A., and Belozerskii, M. A. (1994) Anionic trypsin inhibitors from dry buckwheat seeds∶ isolation, specificity of action, and effect on growth of micromycetes. Biochemistry (Moscow) 59, 739–743.
Urwin, P. E., Atkinson, H. J., Waller, D. A., and McPherson, M. J. (1995) Engineered oryzacystatin-I expressed in transgenic hairy roots confers resistance to Globodera pallida. Plant J. 8, 121–131.
Keen, N. T., Williams, P. H., and Walker, J. C. (1967) Protease of Pseudomonas lachrymans in relation to cucumber angular leaf spot. Phytopathology 57, 263–271.
Hislop, E. C., Paver, J. L., and Keon, J. P. R. (1982) An acid protease produced by Monilinia fructigena in vitro and in infected apple fruits, and its possible role in pathogenesis. J. Gen. Microbiol. 128, 799–807.
Robertsen, B. (1984) An alkaline extracellular protease produced by Cladosporium cucumerinum and its possible importance in the development of scab disease of cucumber seedlings. Physiol. Plant Pathol. 24, 83–92.
Bashan, Y., Okon, Y., and Henis, Y. (1986) A possible role for proteases and deaminases in the development of the symptoms of bacterial speck disease in tomato caused by Pseudomonas syringae pv. tomato. Physiol. Mol. Plant Pathol. 28, 15–31.
Roby, D., Toppan, A., and Esquerré-Tugayé, M. T. (1987) Cell surfaces in plant microorganisms interactions. VIII. Increased proteinase inhibitor activity in melon plants in response to infection by Colletotrichum lagenarium or to treatment with an elicitor fraction from this fungus. Physiol. Mol. Plant Pathol. 30, 453–460.
Tang, J. L., Gough, C. L., Barber, C. E., Dow, J. M., and Daniels M. J. (1987) Molecular cloning of protease gene(s) from Xanthomonas campestris pv. campestris∶ expression in Escherichia coli and role in pathogenicity. Mol. Gen. Genet. 210, 443–448.
Ball, A. M., Ashby, A. M., Daniels, M. J., Ingram, D. S., and Johnstone, K. (1991) Evidence for the requirement of extracellular protease in the pathogenic interaction of Pyrenopeziza brassicae with oilseed rape. Physiol. Mol. Plant Pathol. 38, 147–161.
Dow, J. M., Clarke, B. R., Milligan, D. E., Tang, J. L., and Daniels, M. J. (1990) Extracellular proteases from Xanthomonas campestris pv. campestris, the black rot pathogen. Appl. Environ. Microbiol. 56, 2994–2998.
Dow, J. M., Fan, M. J., Newman, M.-A., and Daniels, M. J. (1993) Differential expression of conserved protease genes in crucifer-attacking pathovars of Xanthomonas campestris. Appl. Environ. Microbiol. 59, 3996–4003.
Bonadé-Bottino, M. (1993) Défense du colza contre les insectes phytophages déprédateurs∶ étude d’une stratégic basée sur l’expression d’inhibiteurs de protéases dans la plante. Ph.D. Thesis, Université de Paris-Sud, Centre d’Orsay.
Leplé, J.-C., Bonadé-Bottino, M., Augustin, S., Delplanque, A., Dumanois, V., Pilate, G., 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.
Benchekroun, A., Michaud, D., Nguyen-Quoc, B., Overney, S., Desjardins, Y., and Yelle, S. (1995) Synthesis of active oryzacystatin I in transgenic potato plants. Plant Cell Rep. 14, 585–588.
Hosoyama, H., Irie, K., Abe, K., and Arai, S. (1994) Oryzacystatin exogenously introduced into protoplasts and regeneration of transgenic rice. Biosci. Biotechnol. Biochem. 58, 1500–1505.
Hilder, V. A., Gatehouse, A. M. R., Sheerman, S. E., Barker, R. F., and Boulter, D. (1987) A novel mechanism of insect resistance engineered into tobacco. Nature 330, 160–163.
Johnson, R., Narvaez, J., An, G., and Ryan, C. A. (1989) Expression of proteinase inhibitors I and II in transgenic potato plants∶ effects on natural defense against Manduca sexta larvae. Proc. Natl. Acad. Sci. USA 86, 9871–9875.
Masoud, S. A., Johnson, L. B., White, F. F., and Reeck, G. R. (1993) Expression of a cysteine proteinase inhibitor (oryzacystatin-I) in transgenic tobacco plants. Plant Mol. Biol. 21, 655–663.
Sloane, B. F., Roxhin, J., Lah, T. T., Day, N. A., Buck, M., Ryan, R. E., Crissman, J. D., and Honn, K. V. (1988) Tumor cathepsin B and its endogenous inhibitors in metastasis. Adv. Exp. Med. Biol. 233P, 259–268.
Mort, J. S., Recklies, A. D., and Poole, A. R. (1984) Extracellular presence of the lysosomal proteinase cathepsin B in rheumatoid synovium and its activity at neutral pH. Arthritis Rheum. 27, 509–515.
Trabandt, A., Gay, R. E., Fassbender, H.-G., and Gay, S. (1991) Cathepsin B in synovial cells at the site of joint destruction in rheumatoid arthritis. Arthritis Rheum. 34, 1444–1451.
Eriksson, S., Janciauskiene, S., and Lannfelt, L. (1995) αl-antichymotrypsin regulates Alzheimer β-amyloid peptide fibril formation. Proc. Natl. Acad. Sci. U.S.A. 92, 2313–2317.
Chapman, H. A., Jr., and Stone, O. L. (1984) Comparison of live human neutrophil and alveolar macrophage elastolytic activity in vitro. Relative resistance of macrophage elastolytic activity to serum and alveolar proteinase inhibitors. J. Clin. Invest. 74, 1693–1700.
Steer, M. L., Meldonlesi, J., and Figarella, C. (1984) Pancreatitis. The role of lysosomes. Dig. Dis. Sci. 29, 934–938.
Kaplan, A. H., Zack, J. A., Knigge, M., Paul, D. A., Kempf, D. J., Norbeck, D. W., and Swanstrom, R. (1993) Partial inhibition of the human immunodeficiency virus type 1 results in aberrant virus assembly and the formation of noninfectious particles. J. Virol. 67, 4050–4055.
Rosé, J. R., Babà, L. M., and Craik, C. S. (1995) Defining the level of human immunodeficiency virus Type I (HIV-1) protease activity required for HIV-1 particle maturation and infectivity. J. Virol. 69, 2751–2758.
Björck, L., Akesson, P., Bohus, M., Trojnar, J., Abrahamson, M., Olafsson, I., and Grubb, A. (1989) Bacterial growth blocked by a synthetic peptide based on the structure of a human proteinase inhibitor. Nature 337, 385–386.
Martin, L. N., Soike, K. F., Murphey-Corb, M., Bohm, R. P., Roberts, E. D., Kakuk, T. K., Thaisrivongs, S., Vidmar, T. J., Ruwart, M. J., Davio, S. R., and Tarpley, W. G. (1994) Effects of U-75875, a peptidomimetic inhibitor of retroviral proteases, on simian immunodeficiency virus infection in rhesus monkeys. Antimicrob. Agents Chemother. 38, 1277–1283.
Korant, B. D., Towatari, T., Ivanoff, L., Petteway, S., Jr., Brzin, J., Lenarcic, B., and Turk, V. (1986) Viral therapy∶ prospects for protease inhibitors. J. Cell. Biochem. 32, 91–95.
Wlodawer, A. and Erickson, J. (1993) Structure-based inhibitors of HIV-1 protease. Annu. Rev. Biochem. 62, 543–585.
Richards, A. D., Roberts, R., Dunn, B. M., Graves, M. C, and Kay, J. (1989) Effective blocking of HIV-1 proteinase activity by characteristic inhibitors of aspartic proteinases. FEBS Lett. 247, 113–117.
Abe, K., Hiroto, K., and Arai, S. (1987) Purification and characterization of a rice cysteine proteinase inhibitor. Agric. Biol. Chem. 51, 2763–2768.
Barrett, A. J. (1987) The cystatins∶ a new class of peptidase inhibitors. Trends Biochem. Sci. 12, 193–196.
Liang, C., Brookhart, G., Feng, G. H., Reeck, G. R., and Kramer, K. J. (1991) Inhibition of digestive proteinase of stored grain Coleoptera by oryzacystatin, a cysteine proteinase inhibitor from rice seeds. FEBS Lett. 278, 139–142.
Chen, M.-S., Johnson, B., Wen, L., Muthukrishnan, S., Kramer, K. J., Morgan, T. D., and Reeck, G. R. (1992) Rice cystatin∶ bacterial expression, purification, cysteine proteinase inhibitory activity, and insect growth suppressing activity of a truncated form of the protein. Protein Expres. Purif. 3, 41–49.
Michaud, D., Nguyen-Quoc, B., and Yelle, S. (1993) Selective inactivation of Colorado potato beetle cathepsin H by oryzacystatins I and II. FEBS Lett. 331, 173–176.
Michaud, D., Bernier-Vadnais, N., Overney, S., and Yelle, S. (1995) Constitutive expression of digestive cysteine proteinase forms during development of the Colorado potato beetle, Leptinotarsa decemlineata Say (Coleoptera∶ Chrysomelidae). Insect Biochem. Mol. Biol., 25, 1041–1048.
Kondo, H., Ijiri, S., Abe, K., Maeda, H., and Arai, S. (1992) Inhibitory effect of oryzacystatins and a truncation mutant on the replication of poliovirus in infected Vero cells. FEBS Lett. 299, 48–50.
Abe, K., Emori, Y., Kondo, H., Arai, S., and Suzuki, K. (1988) The NH2-terminal 21 amino acid residues are not essential for the papain-inhibitory activity of oryzacystatin, a member of the cystatin superfamily. Expression of oryzacystatin cDNA and its truncated fragments in Escherichia coli. J. Biol. Chem. 263, 7655–7659.
Draper, J., Scott, R., and Hamil, J. (1988) Transformation of dicotyledonous plant cells using the Ti plasmid of Agrobacterium tumefaciens and the Ri plasmid of A. rhizogenes, in Plant Genetic Transformation and Gene Expression. A Laboratory Manual (Draper, J., Scott, R., Armitage, P., and Walden, R., eds.), Blackwell Scientific, London, pp. 69–160.
Laemmli, U. K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685.
Smith, B. J. (1984) SDS polyacrylamide gel electrophoresis of proteins, in Methods in Molecular Biology, vol. 1∶ Proteins (Walker, J. M., ed.), Humana, Clifton, NJ, pp. 41–55.
Wenzler, H., Mignery, G., May, G., and Park, W. (1989) A rapid and efficient transformation method for the production of large numbers of transgenic potato plants. Plant Sci. 63, 79–85.
Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989) Molecular cloning∶ A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.
Michaud, D. and Asselin, A. (1995) Review. Application to plant proteins of gel electrophoretic methods. J. Chromatogr. A 698, 263–279.
Bradford, M. M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein using the principle of protein-dye binding. Anal. Biochem. 72, 248–254.
Lowry, O. H., Rosebrough, N. J., Farr, A. L., and Randall, R. J. (1951) Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193, 265–275.
Sarath, G., de la Motte, R. S., and Wagner, F. W. (1989) Protease assay methods, in Proteolytic Enzymes∶ A Practical Approach (Beynon, R. J. and Bond, J. S., eds.), IRL, New York, pp. 25–55.
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.
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. Molecular cloning expression and biochemical studies on oryzacystatin II. J. Biol. Chem. 265, 15832–15837.
Michaud, D., Nguyen-Quoc, B., and Yelle, S. (1994) Production of oryzacystatins I and II in Escherichia coli using the glutathione 5-transferase gene fusion system. Biotechnol. Prog. 10, 155–159.
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Michaud, D., Vrain, T.C. (1998). Expression of Recombinant Proteinase Inhibitors in Plants. In: Cunningham, C., Porter, A.J.R. (eds) Recombinant Proteins from Plants. Methods in Biotechnology, vol 3. Humana Press. https://doi.org/10.1007/978-1-60327-260-5_5
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