FIG 8.

Effects of DAR mutations on interaction between terminal sequences of ZIKV RNA. (A) Schematic representation of RNA molecules used for binding assays. The structures in the 5ʹ RNA and 3ʹ RNA are indicated; the 5ʹ-3ʹ RNA complex is formed by hybridization between the cyclization sequences when the two RNAs are incubated together. (B) RNA binding assay of the DAR mutants. The 3ʹ RNA was incubated at different ratios with WT and DAR mutant 5ʹ RNAs, and the formation of RNA complexes was then analyzed by electrophoresis using a native polyacrylamide gel. The percentages of the bound 3ʹ RNAs, which were calculated by quantification of the band intensities of the 5ʹ-3ʹ RNA complex and the free 3ʹ RNA using the ImageJ software, are shown below the gels.