Genetic transformation of plants to constitutively express virus coat protein genes has been used to confer protection against 5 different plant viruses. Recently, we completed the first field tests with tomato plants and documented field level production against tobacco mosaic virus (TMV). However, the cellular and molecular bases for the engineered protection are not known. Recently, we demonstrated that protection probably blocks an early stage in virus infection that prevents uncoating of the virus. A second manifestation of protection is that systemic movement of virus is slowed in these transgenic plants. We have also conclusively demonstrated that the capsid protein (rather than it's mRNA) is responsible for conferring protection (Powell Abel et al., manuscript in preparation). We now propose to carry out in vivo and in vitro experiments to: (1) determine the role of specific amino acid sequences or protein structures in the protection reaction by producing mutants that affect the sequence and length of the coat protein; (2) determine the amino acid sequences (or protein domains) that confer strain specificity to the protection reaction by producing chimeric proteins comprised of segments of two strains that do not cross-protect against each other (i.e., the common strain and legume strains of (TMV); (3) characterize the protection against systemic virus movement and so determine the role of specific amino acid sequences in such protection. These experiments will lead to a more complete understanding of genetically engineered (cross) protection, and enable the formulation of rational approaches to improve and/or extend the protection.

National Institute of Health (NIH)
National Institute of Allergy and Infectious Diseases (NIAID)
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Virology Study Section (VR)
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Washington University
Schools of Arts and Sciences
Saint Louis
United States
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Bendahmane, Mohammed; Chen, Iju; Asurmendi, Sebastian et al. (2007) Coat protein-mediated resistance to TMV infection of Nicotiana tabacum involves multiple modes of interference by coat protein. Virology 366:107-16
Asurmendi, S; Berg, R H; Smith, T J et al. (2007) Aggregation of TMV CP plays a role in CP functions and in coat-protein-mediated resistance. Virology 366:98-106
Bazzini, A A; Asurmendi, S; Hopp, H E et al. (2006) Tobacco mosaic virus (TMV) and potato virus X (PVX) coat proteins confer heterologous interference to PVX and TMV infection, respectively. J Gen Virol 87:1005-12
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Asurmendi, S; Berg, R H; Koo, J C et al. (2004) Coat protein regulates formation of replication complexes during tobacco mosaic virus infection. Proc Natl Acad Sci U S A 101:1415-20
Koo, Ja Choon; Asurmendi, Sebastian; Bick, Jennifer et al. (2004) Ecdysone agonist-inducible expression of a coat protein gene from tobacco mosaic virus confers viral resistance in transgenic Arabidopsis. Plant J 37:439-48
Bendahmane, Mohammed; Szecsi, Judit; Chen, Iju et al. (2002) Characterization of mutant tobacco mosaic virus coat protein that interferes with virus cell-to-cell movement. Proc Natl Acad Sci U S A 99:3645-50
Staczek, J; Bendahmane, M; Gilleland, L B et al. (2000) Immunization with a chimeric tobacco mosaic virus containing an epitope of outer membrane protein F of Pseudomonas aeruginosa provides protection against challenge with P. aeruginosa. Vaccine 18:2266-74
Opalka, N; Tihova, M; Brugidou, C et al. (2000) Structure of native and expanded sobemoviruses by electron cryo-microscopy and image reconstruction. J Mol Biol 303:197-211

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