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A mammalian 2-5A system functions as an antiviral pathway in transgenic plants

Author:
Mitra, A., Higgins, D.W., Langenberg, W.G., Nie, H., SenGupta, D.N., Silverman, R.H.
Source:
Proceedings of the National Academy of Sciences of the United States of America 1996 v.93 no.13 pp. 6780-6785
ISSN:
0027-8424
Subject:
Nicotiana tabacum, transgenic plants, gene transfer, complementary DNA, ribonucleases, nucleotidyltransferases, humans, gene expression, messenger RNA, enzyme activity, disease resistance, Tobacco mosaic virus, Tobacco etch virus, Alfalfa mosaic virus, leaves
Abstract:
Resistance to virus infections in higher vertebrates is mediated in part through catalysis of RNA decay by the interferon-regulated 2-5A system. A functional 2-5A system requires two enzymes, a 2-5A synthetase that produces 5'-phosphorylated, 2',5'-linked oligoadenylates (2-5A) in response to double-stranded RNA, and the 2-5A-dependent RNase L. We have coexpressed these human enzymes in transgenic tobacco plants by using a single plasmid containing the cDNAs for both human RNase L and a low molecular weight form of human 2-5A synthetase under control of different, constitutive promoters. Expression of the human cDNAs in the transgenic plants was demonstrated from Northern blots, by specific enzyme assays, and by immunodetection (for RNase L). Infection of leaves, detached or in planta, of the coexpressing transgenic plants by tobacco mosaic virus, alfalfa mosaic virus, or tobacco etch virus resulted in necrotic lesions. In contrast, leaves expressing 2-5A synthetase or RNase L alone and leaves containing the plasmid vector alone produced typical systemic infections. While alfalfa mosaic virus produced lesions only in the inoculated leaves regardless of the concentration of virus in the inoculum, high, but not low,levels of tobacco etch virus inoculum resulted in escape of virus to uninoculated leaves. Nevertheless, there was a substantial reduction of tobacco etch virus yield as measured by ELISA assay in the coexpressing transgenic plants. These results indicate that expression of a mammalian 2-5A system in plants provides resistance to virus infections.
Agid:
2356095