Figure 4.

Characterization of RNA elements within HCV IRES domain IV important for HCV IRES–RPS5 interaction. (A) Schematic representation of HCV IRES domain IV wild type and mutant constructs used in the study. Mutated regions are shown in red. (B and C) Direct UV cross-linking of increasing concentrations of 40S ribosomal subunits (5 and 10 pmol) with ³²P-labelled wild type or domain IV mutant IRES RNAs (100 fmol). This is a representative image of three independent experiments. (D) Direct UV cross-linking of increasing concentration of recombinant RPS5 (2 and 4 pmol) with wild-type α-³²P HCV IRES or domain IV IRES mutants (100 fmol). This is a representative image of three independent experiments. (E) In vitro translation of wild type and mutant HCV bicistronic RNAs using rabbit reticulocyte lysate (RRL). Graph represents the percentage luciferase activity (% of F/R). Graph represents the average of three independent experiments.