FIG. 6.

Association of 7SL RNA with viral genomic-RNA-containing RNP complexes. (A) Association of A3C molecules with viral cores. NL4-3 Vif was cotransfected with the A3C expression vector. At 48 h posttransfection, virus supernatants were collected and analyzed on stepwise sucrose gradients. Three fractions were harvested and analyzed by immunoblotting with an HIV-positive human serum to detect CAp24 and Pr55Gag, anti-Map17 and anti-NCp7, or with anti-HA to detect A3C-HA. (B) A3C is not associated with the viral RNP complexes containing viral genomic RNA and primer . 293T cells were transfected with NL4-3ΔVif and the A3C-HA expression vector. Purified virions were treated with nonionic detergent (1% Triton X-100) to disrupt the viral cores. Viral lysates were separated on a linear 20 to 70% (wt/vol) sucrose gradient; 12 fractions were collected from the top of each gradient, and A3C-HA and CAp24 were analyzed by immunoblotting. RNAs were extracted from each fraction and analyzed for the presence of viral genomic RNA and . (C) 7SL RNA molecules are associated with viral genomic RNA and other RNP components inside the viral core structure. Fractions 9 to 12 containing peak viral genomic RNAs were analyzed by qRT-PCR to detect 7SL RNA and by immunoblotting to detect NCp7 and Pr55Gag. The error bars indicate standard deviations.