The amino-terminus of the hepatitis C virus (HCV) p7 viroporin and its cleavage from glycoprotein E2-p7 precursor determine specific infectivity and secretion levels of HCV particle types
Fig 5
p7 ATMI mutant viruses modulate levels of particle-associated viral components.
Huh7.5 cells were electroporated with RNAs from parental vs. Jc1 HAHALp7 mutant viruses expressed alone or with wild-type p7 or HAHALp7, as indicated. Analyses were performed at 72h post-electroporation and normalized by percentage of HCV-positive virus producer cells obtained as in Fig 3A. Results obtained with other p7 ATMI mutant viruses are shown in S6 Fig. (A) Levels of E2 in pellets from ultracentrifuged cell supernatants. (B) Ratio of E2 in pellets vs. intracellular E2. (C) Levels of core and HCV RNA present in the pellets of ultracentrifuged cell supernatants as determined by CMIA and RT-qPCR, respectively. (D) Cell supernatants were incubated with 1% Triton X-100 (TX) or left untreated (-) before ultracentrifugation and analysis of E2 present in the pellets analyzed by quantitative western blot. All values are displayed relative to E2, core or RNA values determined in the pellet of supernatants from Jc1 virus-electroporated cells (A-D). Data represent mean values ± SEM. The numbers of experiments performed are indicated below the graphs.