Mutational analysis of the cucumber necrosis virus coat protein gene

A series of frameshift, deletion, and inversion mutations were made in the coat protein (CP) gene of the icosahedralcucumber necrosis tombusvirus (CNV) to investigate the role of the CP protruding (P) domain in the production of virus particles and, also, to investigate the basis for the accumulation of CP deletion derivatives previously reported in plants inoculated with PD(−), a P-domainless CNV CP mutant. P-domainless coat protein subunit could be detected in extracts of CP mutant-infected plants; however, virus-like particles could not, suggesting that the P domain is essential for tombusvirus particle assembly and/or stability. In addition, each of the P-domain mutants analyzed invariably produced coat protein deletion derivatives in infected plants whereas shell domain mutants rarely produced deletion derivatives. Finally, P-domain inversion and deletion mutants accumulated deletion derivatives very rapidly in comparison to P-domain frameshift mutants. Protoplast studies show that PD(−) RNA inoculum does not undergo further deletion in infected protoplasts, suggesting that PD(−) CP deletion derivatives preferentially accumulate in plants because they have a greater capacity for cell-to-cell movement.