Figure 5
NC mutant viruses display wild-type endogenous reverse transcription kinetics when premature reverse transcription is blocked. Results of endogenous reverse transcription assays performed with virus generated either in the absence or presence of RTIs as indicated at the top of the figure. Virus was prepared using sequential DNase I + subtilisin treatment as described (Figure 1, right). At each time point indicated, a sample of virus was taken and the vDNA was isolated and quantitated. Values were then divided by the number of genomes present at the start of every endogenous reverse transcription reaction to display the quantity of vDNA as a fraction of available genomes. Panels A, C, and E show endogenous reverse transcription time courses using viruses (WT, NCH23C, NCH44C, respectively) prepared without RTIs while panels B, D, and F show time courses with viruses (WT, NCH23C, NCH44C, respectively) prepared in the presence of RTIs. These results are from a representative experiment. The legend for the vDNA species measured is indicated at the bottom of panel A and the bottom of Figure 3 shows schematics of the pertinent vDNA target sites.