Protein Mismatches Caused by Reassortment Influence Functions of the Reovirus Capsid

Following attachment to host receptors via σ1, reovirus particles are endocytosed and disassembled to generate infectious subvirion particles (ISVPs). ISVPs undergo conformational changes to form ISVP*, releasing σ1 and membrane-targeting peptides from the viral μ1 protein. ISVP* formation is required for delivery of the viral core into the cytoplasm for replication. We characterized the properties of T3D^F/T3D^CS1, an S1 gene monoreassortant between two laboratory isolates of prototype reovirus strain T3D: T3D^F and T3D^C T3D^F/T3D^CS1 is poorly infectious. This deficiency is a consequence of inefficient encapsidation of S1-encoded σ1 on T3D^F/T3D^CS1 virions. Additionally, compared to T3D^F, T3D^F/T3D^CS1 undergoes ISVP-to-ISVP* conversion more readily, revealing an unexpected role for σ1 in regulating ISVP* formation. The σ1 protein is held within turrets formed by the λ2 protein. To test if the altered properties of T3D^F/T3D^CS1 are due to a mismatch between σ1 and λ2 proteins from T3D^F and T3D^C, properties of T3D^F/T3D^CL2 and T3D^F/T3D^CS1L2, which express a T3D^C-derived λ2, were compared. The presence of T3D^C λ2 allowed more efficient σ1 incorporation, producing particles that exhibit T3D^F-like infectivity. Compared to T3D^F, T3D^F/T3D^CL2 prematurely converts to ISVP*, uncovering a role for λ2 in regulating ISVP* formation. Importantly, a virus with matching σ1 and λ2 displayed a more regulated conversion to ISVP* than either T3D^F/T3D^CS1 or T3D^F/T3D^CL2. In addition to identifying new regulators of ISVP* formation, our results highlight that protein mismatches produced by reassortment can alter virus assembly and thereby influence subsequent functions of the virus capsid.IMPORTANCE Cells coinfected with viruses that possess a multipartite or segmented genome reassort to produce progeny viruses that contain a combination of gene segments from each parent. Reassortment places new pairs of genes together, generating viruses in which mismatched proteins must function together. To test if such forced pairing of proteins that form the virus shell or capsid alters the function of the particle, we investigated properties of reovirus variants in which the σ1 attachment protein and the λ2 protein that anchors σ1 on the particle are mismatched. Our studies demonstrate that a σ1-λ2 mismatch produces particles with lower levels of encapsidated σ1, consequently decreasing virus attachment and infectivity. The mismatch between σ1 and λ2 also altered the capacity of the viral capsid to undergo conformational changes required for cell entry. These studies reveal new functions of reovirus capsid proteins and illuminate both predictable and novel implications of reassortment.