The Interface between Methyltransferase and Polymerase of NS5 Is Essential for Flavivirus Replication
Figure 5
Characteristics and sequencing analysis of recovered viruses from hydrophobic network and GTR sequence mutations of NS5 in JEV.
(A) Comparison of plaque morphology changes of JEV between representative NS5 mutations and their corresponding passaged viruses. JEV NS5 P113D and F467D viruses were serially passaged in BHK-21 cells, and the plaques at passage 0 (P0) and passage 5 (P5) were compared with that of WT. (B) Summary of sequencing results for all recovered viruses from P5. (C) Crystal structure of JEV NS5 showing the key residues in the MTase-RdRP interface and the recovered secondary compensatory mutations. A combination of cartoon and surface representations was used with MTase in cyan, RdRP palm in grey, thumb in light blue, index finger in green, middle finger in orange, ring finger in yellow, pinky finger in light red, N-terminal extension of RdRP in pink, priming loop in purple, and signature sequence SGDD in magenta. Six hydrophobic residues and the GTR sequence are shown as small spheres that are represented and then labeled at the top left corner. The recovered mutations are shown as large spheres with the same color of its original mutations.