Fig. 1: Ago2 occupancy at Site-1 protects HCV RNA from degradation and crystal structure of the protective complex.

a Cartoon schematic of Ago2:miR-122 complex (left). Schematic of the HCV 5’ UTR with Ago2-miR-122 complexes bound to miR-122 binding Site-1 and Site-2 at the 5’ end of the genome, upstream of the IRES (right). Stem-loop 1 (SL1) is embedded within miR-122 Site-1. b Degradation of HCV 5’ UTR wild type (WT) RNA by XRN1 in the presence or absence of Ago2:miR-122 over time. Data from 3 independent replicates with error bars representing SEM. c Degradation of HCV 5’ UTR RNAs by XRN1 with mutations that disrupt miR-122 binding at Site-1 (MUT1) or Site-2 (MUT2) compared to WT 5’ UTR data from 1B. Data from 3 independent replicates with error bars representing SEM. d Degradation of HCV 5’ UTR ∆SL1 RNA and HCV 5’ UTR ∆SL1 MUT2 RNA by XRN1 compared to WT data from 1B. Data from 3 independent replicates with error bars representing SEM. e Relative abundance over 5 days of viral RNA after electroporation into Huh-7 cells. Mutations (WT, MUT1, MUT2, ∆SL1, NS5B GDD > AAG) indicated. Data from 3 independent replicates with error bars representing SEM. f Crystal structure of Ago2:miR-122 bound to HCV Site-1 RNA. Ago domains are coloured from grey (N-terminal), through olive (PAZ and MID), to gold (PIWI). miR-122 in red, HCV RNA in blue. Stem-loop 1 (SL1) extends from the center of the complex. g Front, top and side views with Ago2 in surface representation (grey).