FIG. 5.
Placement of tRNA3Lys on HIV-1 1-331 RNA by GagΔp6. (A) 32P-labeled tRNA3Lys was incubated with retroviral transcript and GagΔp6, and the reactions were analyzed as described in Materials and Methods. Lanes: 1, tRNA alone; 2, tRNA plus HaSV 34-378 plus 6 μg of GagΔp6; 3, tRNA plus HIV-1 1-331 plus 3 μg of GagΔp6; 4, tRNA plus HIV-1 1-331 plus 6 μg of GagΔp6; 5, tRNA plus HIV-1 1-331. (B) Unlabeled tRNA3Lys was annealed to the PBS on HIV-1 1-331 RNA by incubation as indicated. The mixtures were deproteinized as described in Materials and Methods and then tested for the presence of primer on the HIV-1 template RNA by addition of reverse transcriptase and dNTPs, including [α-32P]dCTP. The reactions were analyzed as described in Materials and Methods. Size of the labeled DNA product was determined ± 10 nt compared with labeled DNAs of known sizes (data not shown). Lanes: 1, tRNA plus 6 μg of GagΔp6; 2, 6 μg of GagΔp6 plus HIV-1 1-331; 3, tRNA plus HIV-1 1-331; 4, tRNA plus 6 μg of GagΔp6 plus HIV-1 1-331.