FIG. 3.

The effect of HIV-1 Pol multimerization-defective mutants on RNA dimer maturation. (A) Schematic representation of HIV-1 proviral DNA constructs containing amino acid substitutions at the indicated sites that prevent RT dimerization is given: L234A (L) and W401A (W) or IN multimerization V260E (V). Further combinations of these substitutions were also generated. (B) Western blot analysis was performed on purified virions. Proteins were resolved by 10% Tris-glycine SDS-PAGE and then probed with pooled sera from HIV-1-infected individuals. (C) RNA dimer analysis was performed on WT, PR(−) (which contains an inactive protease), W401A, and V260E single mutants; the W401A and V260E double mutant (WV); and the L234A, W401A, and V260E triple mutant (LWV). Virion RNA was resuspended in RNA dimerization buffer and heat denatured for 10 min at the indicated temperatures. Dimers and monomers were electrophoresed in a 1% native agarose gel and probed with an HIV-1-specific riboprobe.