Env adaptations increase RT packaging, accelerate reverse transcription, and reduce G-to-A mutation levels.
(A) Representative immunoblots of the indicated proteins in viral particles and producer cells from one experiment. (B) Relative RT packaging into viral particles produced in SupT11-A3G cells. RT packaging levels were quantified based on band intensity and normalized by each p24 of the virions (mean +/- SD of 3 biologically independent experiments). (C) Relative RT activity in viral particles produced in SupT11-A3G cells. RT activity was measured for each viral lysate normalized to p24 levels (mean +/- SD of 3 biologically independent experiments). (D to G) Kinetics of early RT, late RT, 2-LTR circle, and proviral DNA during infection of CEM-GFP cells using viruses originally produced in SupT11-A3G cells (mean +/- SD of 3 biologically independent experiments). (H) G-to-A mutation loads of high-fidelity, high-temperature pol amplicons from CEM-GFP cells infected with the indicated viruses (mean +/- SD of 3 biologically independent experiments with a minimum of 10 sequences or 5,640 bp per condition). Statistical comparisons for data in panels B-H were done using Student’s t test (p-values above each panel in comparison to Vif-null HIV-1; *: p<0.05, **: p<0.01, ***: p<0.001).