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. Author manuscript; available in PMC: 2022 Jun 16.
Published in final edited form as: J Mol Biol. 2021 Oct 16;433(24):167306. doi: 10.1016/j.jmb.2021.167306

Figure 10. Amino acids 144 in combination with 44 and 45 in rhA3C enable HIV restriction ability.

Figure 10.

(A) Infectivity was measured by β-galactosidase expression driven by the HIV-1 5΄LTR from TZM-bl cells infected with VSV-G pseudotyped HIV ΔVif ΔEnv that was produced in the absence or presence of 3xHA tagged hA3C S188I, rhA3C WT, and rhA3C mutants S144A and Q44R/H45R/S144A. Results normalized to the no A3 condition are shown with error bars representing the Standard Deviation of the mean calculated from three independent experiments. (B) Immunoblotting for the HA tag was used to detect A3C enzymes expressed in cells and encapsidated into HIV ΔVif ΔEnv pseudotyped virions. The cell lysate and virion loading controls were α-tubulin and p24, respectively. (C) The relative amount of proviral DNA integration in infected HEK293T cells in the presence of hA3C S188I, rhA3C WT, and rhA3C mutants Q44R/H45R/S144A and Q44R/H45R/M115N in comparison to the No A3 condition was determined by qPCR. Error bars represent the standard deviation of the mean calculated from two independent experiments.