The structure of a 12-segmented dsRNA reovirus: New insights into capsid stabilization and organization.

Infecting a wide range of hosts, members of Reovirales (formerly Reoviridae) consist of a genome with different numbers of segmented double stranded RNAs (dsRNA) encapsulated by a proteinaceous shell and carry out genome replication and transcription inside the virion. Several cryo-electron microscopy (cryo-EM) structures of reoviruses with 9, 10 or 11 segmented dsRNA genomes have revealed insights into genome arrangement and transcription. However, the structure and genome arrangement of 12-segmented Reovirales members remain poorly understood. Using cryo-EM, we determined the structure of mud crab reovirus (MCRV), a 12-segmented dsRNA virus that is a putative member of Reovirales in the non-turreted Sedoreoviridae family, to near-atomic resolutions with icosahedral symmetry (3.1 Å) and without imposing icosahedral symmetry (3.4 Å). These structures revealed the organization of the major capsid proteins in two layers: an outer T = 13 layer consisting of VP12 trimers and unique VP11 clamps, and an inner T = 1 layer consisting of VP3 dimers. Additionally, ten RNA dependent RNA polymerases (RdRp) were well resolved just below the VP3 layer but were offset from the 5-fold axes and arranged with D₅ symmetry, which has not previously been seen in other members of Reovirales. The N-termini of VP3 were shown to adopt four unique conformations; two of which anchor the RdRps, while the other two conformations are likely involved in genome organization and capsid stability. Taken together, these structures provide a new level of understanding for capsid stabilization and genome organization of segmented dsRNA viruses. Author summary: Mud crab reovirus (MCRV), a putative member of the Sedoreoviridae family in Reovirales, infects an economically important crab, Scylla serrata, and has resulted in serious economic loss for the crab aquaculture. In this study, we report the structure of MCRV, which has allowed us to examine the detailed structural elements that might be responsible for capsid stability, as well as those involved in genome packaging and RdRp organization. The arrangement of viral polymerases inside the capsid differ from what has been seen in other viruses of the same order, as ten of the vertices are occupied by RdRp in a symmetrical organization not seen elsewhere. In addition, the VP11 protein is analogous to the "clamp" protein of turreted reovirads. [ABSTRACT FROM AUTHOR]