Optimum length and flexibility of reovirus attachment protein σ1 are required for efficient viral infection.

Bokiej M, Ogden KM, Ikizler M, Reiter DM, Stehle T, Dermody TS
J Virol. 2012 86 (19): 10270-80

PMID: 22811534 · PMCID: PMC3457286 · DOI:10.1128/JVI.01338-12

Reovirus attachment protein σ1 is an elongated trimer with head-and-tail morphology that engages cell-surface carbohydrate and junctional adhesion molecule A (JAM-A). The σ1 protein is comprised of three domains partitioned by two flexible linkers termed interdomain regions (IDRs). To determine the importance of σ1 length and flexibility at different stages of reovirus infection, we generated viruses with mutant σ1 molecules of altered length and flexibility and tested these viruses for the capacity to bind the cell surface, internalize, uncoat, induce protein synthesis, assemble, and replicate. We reduced the length of the α-helical σ1 tail to engineer mutants L1 and L2 and deleted midpoint and head-proximal σ1 IDRs to generate ΔIDR1 and ΔIDR2 mutant viruses, respectively. Decreasing length or flexibility of σ1 resulted in delayed reovirus infection and reduced viral titers. L1, L2, and ΔIDR1 viruses but not ΔIDR2 virus displayed reduced cell attachment, but altering σ1 length or flexibility did not diminish the efficiency of virion internalization. Replication of ΔIDR2 virus was hindered at a postdisassembly step. Differences between wild-type and σ1 mutant viruses were not attributable to alterations in σ1 folding, as determined by experiments assessing engagement of cell-surface carbohydrate and JAM-A by the length and IDR mutant viruses. However, ΔIDR1 virus harbored substantially less σ1 on the outer capsid. Taken together, these data suggest that σ1 length is required for reovirus binding to cells. In contrast, IDR1 is required for stable σ1 encapsidation, and IDR2 is required for a postuncoating replication step. Thus, the structural architecture of σ1 is required for efficient reovirus infection of host cells.

MeSH Terms (21)

Animals Capsid Capsid Proteins Carbohydrates Cell Adhesion Molecules Cell Line Cell Membrane Fibroblasts Fluorescent Antibody Technique, Indirect Gene Deletion Gene Expression Regulation, Viral Genome, Viral HeLa Cells Humans Mice Mutation Protein Binding Protein Folding Receptors, Cell Surface Reoviridae Virus Replication

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