Nature Biotechnology
14, 1566 - 1569 (1996)
doi:10.1038/nbt1196-1566
Virus-induced cell death in plants expressing the mammalian 2',5' oligoadenylate systemToshiya Ogawa1, Tamaki Hori1
& Isao Ishida1, *
1Central Laboratories for Key Technology, 1-13-5, Fukuura, Kanazawa-ku Yokohama-shi, Kanagawa236, Japan.
*e-mail: i-ishida@kirin.co.jp The major components of the 2-5A system, responsible for the mammalian interferon-induced antiviral response, are the 2',5' oligoadenylate synthetase (2-5Aase) and 2',5' oligoadenylate (2-5A) dependent ribonuclease (RNase L). Transgenic tobacco plants expressing these two enzyme activities were produced by crossing the transgenic plants expressing RNase L with those expressing 2-5Aase. The double transgenic plants showed complete resistance against cucumber mosaic virus (CMV), infection with necrotic spots only forming on the virus-inoculated leaf. On the other hand, although plants inoculated with potato virus Y (PVY) formed necrotic spots on the inoculated leaf and virus amplification could not be detected, all plants died within 20 days of inoculation. The transgenic tobacco plants expressing either 2-5Aase or RNase L activity showed typical disease symptoms with CMV- or PVY-inoculation. These results suggest that the introduced 2-5A system is activated in tobacco cells by dsRNA, the replicating intermediates of RNA viruses, leading to death of the host cells, which has not been observed in mammalian cells.
REFERENCES
- Fitchen, J.H. and Beachy, R.N. 1993. Genetically engineered protection against viruses in transgenic plants. Annu. Rev. Microbiol. 47: 739−763. | Article | PubMed | ISI | ChemPort |
- Lodge, J.K., Kaniewsky, W.K., and Turner, N.E. 1994. Broad-spectrum virus resistance in transgenic plants expressing pokeweed antiviral protein. Proc. Natl. Acad. Sci. USA 90: 7089−7093.
- Truve, E., Aaspollu, A., Honkanen, J., Puska, R., Mehto, M., Hassi, A., et al. 1993. Transgenic potato plants expressing mammalian 2'-5' oligoadenylate synthetase are protected from potato virus X infection under field conditions. Bio/Technology 11: 1048−1052. | PubMed | ISI | ChemPort |
- Watanabe, Y., Ogawa, T., Takahashi, H., Ishida, I., Takeuchi, Y., Yamamoto, M., and Okada, Y. 1995. Resistance against multiple plant viruses in plants mediated by a double stranded-RNA specific ribonuclease. FEBS Letters 372: 165−168. | Article | PubMed | ISI | ChemPort |
- Lawson, C., Kaniewski, W., Haley, L., Rozman, R., Newell, C., Sanders, P., and Turner, N.E. 1990. Engineering resistance to mixed virus infection in a commercial potato cultiver: Resistance to potato virus X and potato virus Y in transgenic Russet Burbank. Bio/Technology 8: 127−134 | PubMed | ISI | ChemPort |
- Lengyel, P. 1982. Biochemistry of interferons and their action. Annu. Rev. Biochem. 51: 251−282. | Article | PubMed | ISI | ChemPort |
- Benech, P., Mory, Y., and Chebath, J. 1985. Structure of two forms of the inter-feron-induced (2'-5') oligo A synthetase of human cells based on cDNAs and gene sequences. EMBO J. 4: 2249−2256. | PubMed | ISI | ChemPort |
- Zhou, A., Hassel, B.A., and Silverman, R.H. 1993. Expression cloning of 2-5A-dependent RNAase: A uniquely regulated mediator of interferon action. Cell 2: 753−765. | Article |
- Wells, J.A., Swyryd, E.A., and Stark, G.R. 1984. An improved method for purifying 2',5'-oligoadenylate synthetases. J. Biol. Chem. 259: 1363−1370. | PubMed | ISI | ChemPort |
- Silverman, R.H. 1985. Functional analysis of 2-5A-dependent RNase and 2-5A using 2',5'-oligoadenylate-cellulose. Anal. Biochem. 144: 450−460. | PubMed | ISI | ChemPort |
- Okuno, T., Nakayama, M., Yoshida, S., Furusawa, I., and Komiya, T. 1993. Comparative susceptibility of transgenic tobacco plants and protoplasts expressing the coat protein gene of cucumber mosaic virus to infection with virions and RNA. Phytopathology 83: 542−547. | ISI | ChemPort |
- Ohshima, K., Hataya, T., Sano, T., Inoue, A.K., and Shikata, E. 1991. Comparison of biological properties, serological characteristics and amino acid sequences of coat protein between potato virus Y ordinary strain and necrotic strain. Ann. Phytopath. Soc. Japan 57: 615−622. | ChemPort |
- Meshi, T., Meshi, T., and Okada, Y. 1982. Nucleotide sequence and its character of cistron coding for the 30 kilodalton protein of tobacco mosaic virus DM strain. J. Biochem. 91: 1441−1444. | PubMed | ISI | ChemPort |
- Whitham, S., Dinesh-Kumar, S.P., Choi, D., Hehl, R., Corr, C., and Baker, B. 1994. The product of the tobacco mosaic virus resistance gene N: Similarity to toll and the interleukin-1 receptor. Cell 78: 1101−1115. | Article | PubMed | ISI | ChemPort |
- Devash, Y., Reichman, M., Sela, I., Reichenbach, N.L., and Suhadolnik, R.J. 1985. Plant oligoadenylates: enzymatic synthesis, isolation, and biological activities. Biochemistry 24: 593−599. | PubMed | ISI | ChemPort |
- Truve, E., Kelve, M., Aaspollu, A., Kuusksalu, A., Seppanen, P., and Saarma, M. 1994. Principles and background for the construction of transgenic plants displaying multiple virus resistance. Arch. Virol. [suppl] 9: 41−50. | ChemPort |
- Mehdy, M.C. 1994. Active oxygen species in plant defense against pathogens. Plant Pathol. 105: 467−472. | ChemPort |
- Valverde, R.A., Dodds, J.A., and Heick, J. 1986. Double-stranded RNA from plants infected with virus having elongated particles and undivided genomes. Phytopathology 76: 459−465. | ISI | ChemPort |
- Silverman, R.H., Jung, D.D., Nolan-Sorden, N.L., Dieffenbach, C.W., Kedar, V.P., and SenGupta, D.N. 1988. Purification and analysis of murine 2-5A-dependent RNase. J. Biol. Chem. 263: 7336−7341. | PubMed | ISI | ChemPort |
- Puska, R., Seppanen, P., Honkanen, J., Truve, E., Aaspollu, A., Nigul, L., and Saarma, M. 1994. Transgenic plants show protection against RNA viruses. The 4th International Congress of Plant Molecular Biology (Amsterdam), Abstract.
- Mitra, A., Higgins, D.W., Langenberg, W.G., Nie, H., Sengupta, D.N., and Silverman, R. 1996. A mammalian 2-5A system functions as an antiviral pathway in transgenic plants. Proc. Natl. Acad. Sci. USA 93: 6780−6785. | Article | PubMed | ChemPort |
- Towbin, H., Gordon, J., and Staehelin, T. 1979. Electrophoretic transfer of proteins from polyacrylamide gels to nitro cellulose sheets—procedure and some
|
