C. D. Consalvo, P. J. Nichols, E. P. Boyle, P. J. Aruscavage, P. S. Shen, B. L. Bass
In prior studies we showed that the nematode ancestor of Dicer's helicase domain had minimal ATP hydrolysis, translocation and dsRNA binding activity, and in extant C. elegans the RIG-I like receptor (RLR) DRH-1, and the dsRNA binding protein RDE-4, were co-opted to provide these activities. Here we report cleavage-competent cryo-EM structures of the antiviral complex (AVC; DCR-1{middle dot}DRH-1{middle dot}RDE-4), in the absence (3.2[A]) and presence (3.0[A] of ATP. A key feature of both structures is stacking of DCR-1's helicase with the helicases of two DRH-1 molecules, reminiscent of oligomerization of mammalian RLRs. dsRNA threads through all three helicases with a widened major groove at helicase-helicase interfaces and where the conserved Hel2 loop inserts into the major groove. The presence of nucleotide redistributed helicase-helicase and helicase-RNA contacts in the AVC, as for MDA5, albeit specific interactions and remodeling differed. MDA5 and RIG-I RLRs contain an unstructured linker between their CARDs and helicase domain, but the analogous linker in DRH-1 has a short-structured region positioned to interact with DCR-1. These findings provide a structural framework for understanding how DCR-1, DRH-1, and RDE-4 cooperate to cleave viral dsRNA and elucidate unique features required for antiviral defense in different animals.