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Significance Bluetongue virus (BTV) is a double-stranded RNA (dsRNA) virus that often causes significant economic damage to the livestock industry. The RNA-based genome transcription of BTV during infection depends on its RNA-dependent RNA polymerase (RdRp) inside the capsid. Here, we report a structure of BTV RdRp derived from cryo-electron microscopy (cryoEM) and reconstruction methods. The structure reveals how BTV RdRp interacts with associated genomic dsRNA and capsid proteins. Structural comparison between RdRp in the transcriptionally incompetent BTV virion and transcriptionally competent BTV cores provides insights into the highly regulated mechanism of mRNA transcription initiation. This structure of BTV RdRp and our mechanistic findings could now be exploited to design antivirals against the bluetongue disease in the livestock farming industry.

In situ structures of RNA-dependent RNA polymerase inside bluetongue virus before and after uncoating