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. Author manuscript; available in PMC: 2021 Jan 15.
Published in final edited form as: Virology. 2019 Nov 15;540:119–131. doi: 10.1016/j.virol.2019.11.010

Figure. 6. GADD34 inhibits HIV-1 protein expression by viral 5'-UTR TAR RNA-mediated translational inhibition.

Figure. 6.

A) Schematic representation of human IFIT1-promoter-Luc plasmid in which HIV-1 5’-UTR (1-289 nt) and its deletion mutants were cloned downstream of IFIT1 promoter RNA initiation site.

B) 293FT cells were transfected with 0.5 μg of IFIT1-promoter-Luc or indicated IFIT1-promoter-HIV-1- 5’-UTR-Luc chimeric plasmids in the presence or absence of 0.5 μg of GADD34/Stop or GADD34 plasmids for 36h. The effect of GADD34 transfection on the luciferase activity of IFIT1-promoter and the chimeric plasmids is plotted as a percentage of the effect of the control GADD34/Stop plasmid on the luciferase activity. The bar labeled GADD34/Stop represents luciferase activity of each IFIT1-Luc construct in the presence of GADD34/Stop set at 100 percent. C) 293FT cells were transfected with 0.5 μg of IFIT-UTR-1-289 plasmid in the presence of indicated plasmids. Cells were harvested 38 h later for luciferase measurement. The effect of transfection of indicated GADD34 plasmids on the luciferase activity of IFIT-UTR-1-289-Luc plasmid is plotted as a percentage of the effect of the control GADD34/Stop plasmid on the IFIT-UTR-1-289 plasmid- driven luciferase activity. The bar labeled GADD34/Stop represents luciferase activity of the IFIT-UTR-1-289 plasmid in the presence of GADD34/Stop set at 100 percent. D) HIV-1 TAR RNA and Poly A hairpin regions with mutations shown in bold letters. E) 293FT cells were transfected with 0.5 μg of the indicated plasmids in the presence or absence of 0.5 μg of GADD34/Stop or GADD34 plasmids. Cells were harvested 38 h later for luciferase measurement. The effect of GADD34 transfection on the luciferase activity of IFIT1-promoter and the chimeric plasmids is plotted as a percentage of the effect of the control GADD34/Stop plasmid on the luciferase activity. The bar labeled GADD34/Stop represents luciferase activity of each IFIT1-Luc construct in the presence of GADD34/Stop set at 100 percent.