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. 2002 Mar;76(5):2329–2339. doi: 10.1128/jvi.76.5.2329-2339.2002

FIG. 7.

FIG. 7.

Model for RNase H-mediated degradation of the vRNA template during reverse transcription. In the PBS occupancy assay, both the downstream AUG DNA primer and the tRNA primer are extended by RT. Extension of the tRNA primer yields a full-length tRNA-cDNA on the wild-type template (top) but not on the PAS-negative template (bottom). Extension of the tRNA primer triggers RNase H-mediated degradation of the RNA template (arrows and breaks in the processed RNA). The RNase H domain of HIV-1 RT is located 18 nt from the polymerase active site (24, 26). Thus, enhanced pausing of the RT enzyme near template position +154 on the PAS-negative template results in pronounced RNase H cleavage at position +172 (thick arrow). AUG-primed reverse transcription complexes that displace the PBS-bound tRNA primer stop at this position are shown. The HIV-1 RT enzyme covers approximately 8 nt upstream and 22 nt downstream of the polymerization site (34, 44), such that the reversion-based C150U mutation is within the RT complex.