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 6
p7 ATMI mutant viruses induce secretion of viral particles with impaired envelopment.
Huh7.5 cells were electroporated with RNAs from parental vs. Jc1 HAHALp7 or JFH1 HAHALp7 mutant viruses expressed alone or with wild-type p7 or HAHALp7, as indicated. Analyses were performed at 72h post-electroporation. (A) Levels of core and RNA following GNA pull down of cell supernatants, as determined by CMIA and RT-qPCR, respectively, and normalized by the amounts of immuno-precipitated E2. The values are displayed relative to association of core or RNAs to E2 pulled-down from the supernatants of Jc1 virus-electroporated cells. (B) Cell supernatants were digested with proteinase K (+ PK) with (+ TX) or without pre-treatment with Triton X-100 and the residual amounts of core (i.e., lipid membrane-protected core) were determined by CMIA. The values are displayed relative to non-treated conditions. (C) Analysis of density gradients of cell supernatants. Infectivity, core, RNA, and E2 were measured in each fraction and expressed as percentage of the sum of fractions. Examples of raw data of infectivity can be found in S7A–S7C Fig. (D) Infectivity and core of the indicated viruses were measured in pooled fractions of densities of 1.08–1.15 g/ml and represented as data normalized by values obtained with parental viruses. The grey shaded area represents values below the sensitivity threshold of the experiments. Data represent mean values ± SEM.