Incorporation of epitope-tagged viral sigma3 proteins to reovirus virions.
Tagging of viral capsid proteins is a powerful tool to study viral assembly; it also raises the possibility of using viral particles to present exogenous epitopes in vaccination or gene therapy strategies. The ability of reoviruses to induce strong mucosal immune response and their large host range and low pathogenicity in humans are some of the advantages of using reoviruses in such applications. In the present study, the feasibility of introducing foreign epitopes, "tags", to the sigma3 protein, a major component of the reovirus outer capsid, was investigated. Among eight different positions, the amino-terminal end of the protein appeared as the best location to insert exogenous sequences. Additional amino acids at this position do not preclude interaction with the micro1 protein, the other major constituent of the viral outer capsid, but strongly interfere with micro1 to micro1C cleavage. Nevertheless, the tagged sigma3 protein was still incorporated to virions upon recoating of infectious subviral particles to which authentic sigma3 protein was removed by proteolysis, indicating that micro1 cleavage is not a prerequisite for outer capsid assembly. The recently published structure of the sigma3- micro1 complex suggests that the amino-terminally inserted epitope could be exposed at the outer surface of viral particles.
Départment de microbiologie et immunologie, Université de Montréal, Canada.
Etienne
E
Rouault
Guy
G
Lemay
D000595:Amino Acid Sequence; D000818:Animals; D019556:COS Cells; D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism*; D000939:Epitopes / Q000276:immunology*; D006639:Histidine; D015151:Immunoblotting; D007739:L Cells; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016601:RNA-Binding Proteins; D012087:Reoviridae / Q000378:metabolism*; D014162:Transfection; D014771:Virion / Q000276:immunology / Q000378:metabolism*; D019065:Virus Assembly
D036022:Capsid Proteins; D000939:Epitopes; D016601:RNA-Binding Proteins; C055356:mu1 protein, Reovirus; C053179:sigma protein 3, Reovirus; D006639:Histidine