Figure 7

Effect of non-specific RNA-binding peptide on encapsidation and antiviral activity of CD1 mutant H65R. (A) Strucures of constructs P22-H65R, H65R, P22-A3G, and wild-type A3G. (B) Western blotting analysis of lysates of 293T cells co-transfected with the A3G constructs as well as pHDV-EGFP, C-HelpΔVif and pHCMV-G (panel labeled Cell lysate). The cell lysates were also analyzed using anti-tubulin antibody to insure equivalent loading of cell lysate proteins (panel labeled α-tubulin). Incorporation of A3G BiFC constructs into VLPs (panel labeled Viral lysate) was determined by analyzing viral lysates containing 100 ng of p24 CA. (C) Relative cytidine deaminase activity in viral lysates of virions produced in the presence of wild type A3G, P22-A3G, P22-H65R, H65R, or in the absence of A3G. The cytidine deaminase activity of A3G-CY was set to 100%. (D) Relative HDV-EGFP infectivity in the presence of P22-A3G, P22-H65R and H65R constructs. Infectivity of the virions produced was determined by flow cytometry analysis of cells infected with the virions. The proportion of GFP⁺ cells after infection with HDV-EGFP in the absence of A3G was set to 100%. Error bars represent the s.e.m. of three independent experiments.