title: Characterization of the thermosensitive ts453 reovirus mutant: increased dsRNA binding of sigma 3 protein correlates with interferon resistance.
abstract: The mutation harbored by the reovirus ts453 thermosensitive mutant has been assigned to the S4 gene encoding the major outer capsid protein sigma 3. Previous gene sequencing has identified a nonconservative amino acid substitution located near the zinc finger of sigma 3 protein in the mutant. Coexpression in COS cells of the sigma 3 protein presenting this amino acid substitution (N16K), together with the other major capsid protein mu 1, has also revealed an altered interaction between the two proteins; this altered interaction prevents the sigma 3-dependent cleavage of mu 1 to mu 1C. 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 sigma 3 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 sigma 3 nuclear presence. The lack of association with mu 1, and/or the increased dsRNA-binding activity of sigma 3, 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.
authors.affiliation: Département de Microbiologie et Immunologie, Université de Montréal, Québec, Canada.
authors.forename: J
authors.initials: J
authors.lastname: Bergeron
authors.forename: T
authors.initials: T
authors.lastname: Mabrouk
authors.forename: S
authors.initials: S
authors.lastname: Garzon
authors.forename: G
authors.initials: G
authors.lastname: Lemay
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D019556:COS Cells; D002213:Capsid / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D036022:Capsid Proteins; D002460:Cell Line; D004352:Drug Resistance, Microbial; D016899:Interferon-beta / Q000494:pharmacology*; D051379:Mice; D016297:Mutagenesis, Site-Directed; D012330:RNA, Double-Stranded / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D012087:Reoviridae / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D013696:Temperature; D014764:Viral Proteins / Q000096:biosynthesis; D015032:Zinc / Q000378:metabolism
chemical_list: D000998:Antiviral Agents; D036022:Capsid Proteins; D012330:RNA, Double-Stranded; D016601:RNA-Binding Proteins; D014764:Viral Proteins; C053179:sigma protein 3, Reovirus; D016899:Interferon-beta; D015032:Zinc