Retroviral Restriction Factor APOBEC3G Delays the Initiation of DNA Synthesis by HIV-1 Reverse Transcriptase
Figure 5
A3G E259Q inhibition of RT-mediated primer extension from the HIV-1 PBS.
An 18 nt 32P-labeled RNA primer containing a sequence complementary to the HIV-1 PBS was annealed to a 106 nt RNA containing the PBS (sketch). Complete extension of the primer results in a product of 82 nt (sketch). The p/t was used at a concentration of 10 nM. (A) Primer extension by RT (480 nM) in the absence (0∶1) or presence (4∶1, 48∶1) of A3G E259Q relative to the p/t concentration. Reactions were sampled at 5 and 60 min. (B) Quantification of primer extension (%) from gel shown in panel A for 5 min (black bars) and 60 min (grey bars). Primer extension (%) values are averaged from three independent trials. Designations for significant difference from 0∶1 values were p≤0.001 (***), p≤0.01(**), or p≤0.05 (*).