Aminoterminal Amphipathic α-Helix AH1 of Hepatitis C Virus Nonstructural Protein 4B Possesses a Dual Role in RNA Replication and Virus Production
Figure 6
Analysis of pseudorevertants selected with NS4B AH1 mutant K18A.
(A) Selected amino acid changes identified following transfection of a selectable subgenomic replicon harboring the NS4B AH1 K18A mutation (see Table 1) were reengineered into a subgenomic HCV replicon harboring a luciferase reporter gene. In vitro transcribed RNAs were electroporated into Huh-7.5 cells, followed by luciferase activity measurement at 4, 24, 48 and 72 h post-electroporation. Relative light units (RLU) were normalized to values measured at 4 h. Results from two independent experiments were pooled (n = 8 for each construct and time point). For statistical analyses, RLU measured at 48 h were compared to the 48-h values obtained with mutant K18A. *, P<0.001. (B) Structure models of the different AH1 sequences were established by using Swiss-PdbViewer 4.01 software [69] and drawn by using VMD 1.9 software [68].