Characterization of the Thermosensitive ts453 Reovirus Mutant: Increased dsRNA Binding of ς3 Protein Correlates with Interferon Resistance

AbstractThe mutation harbored by the reovirus ts453 thermosensitive mutant has been assigned to the S4 gene encoding the major outer capsid protein ς3. Previous gene sequencing has identified a nonconservative amino acid substitution located near the zinc finger of ς3 protein in the mutant. Coexpression in COS cells of the ς3 protein presenting this amino acid substitution (N16K), together with the other major capsid protein μ1, has also revealed an altered interaction between the two proteins; this altered interaction prevents the ς3-dependent cleavage of μ1 to μ1C. This could explain the lack of outer capsid assembly observed during ts453 virus infection at nonpermissive temperature. In the present study, we pursued the characterization of this mutant ς3 protein. Although the N16K mutation is located close to the zinc finger region, it did not affect the ability of the protein to bind zinc. In contrast, this mutation, as well as mutations within the zinc finger motif itself, can increase the binding of the protein to double-stranded RNA (dsRNA). It also appears that the N16K mutant protein is more efficiently transported to the nucleus than the wild-type protein, an observation consistent with the postulated role of dsRNA binding in ς3 nuclear presence. The lack of association with μ1, and/or the increased dsRNA-binding activity of ς3, could be responsible for a partial resistance of the ts453 virus to interferon treatment and this could have important consequences in the context of protein synthesis regulation during natural reovirus infection