mini-RNP_revision_map&models_SourceData

Maps&amp;Models and Source data related to manuscript: <b>Coupling of polymerase-nucleoprotein-RNA in an influenza virus mini ribonucleoprotein complex</b>.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.