mini-RNP_revision_map&models_SourceData

Maps&Models and Source data related to manuscript: Coupling of polymerase-nucleoprotein-RNA in an influenza virus mini ribonucleoprotein complex.Manuscript AbstractInfluenza virus polymerase complex (FluPol), nucleoprotein (NP) and RNA constitute the ribonucleoprotein complexes (RNPs) that play essential roles in virus replication and transcription. Here we report the cryo-EM structures of influenza virus mini viral RNPs (mini-vRNPs) reconstructed by RNP components in two distinct states at atomic resolution, among which FluPol is either in the inner side (State-In) or at the outer rim (State-Out) of the NP-RNA ring. In both states, the 5¢ and 3¢ termini of vRNA are bound to FluPol as previously reported. One NP (NP-0) contacts PA/PB1 of FluPol, and binds to the double-stranded distal part of the vRNA promoter that projects away from FluPol. The D72-K90 loop which contain an ɑ-helix (residues D72-E81) in NP-0 inserts into the fork of the paired vRNA, and the separated single strands are bound in the RNA binding grooves of NP-0. The RNA binding grooves of the other NPs form a continuous path to sequester the RNA, similar to the common mechanism used by NPs of negative-strand RNA viruses to protect the genome. The interfaces for FluPol dimerization or interactions with Pol II are obstructed in the State-In structure, but are fully exposed in the State-Out structure. These structures dissect the details for the coupling of FluPol, NP and RNA in mini-vRNPs, and suggest a model for the conformational shift of RNP that may occur during the life cycle of the virus.