FIG. 4.

Envelope glycoprotein CDs from diverse lentiviruses interact with PRA1. (A) The Env CDs of SIVmac239, HIV-1 NL4-3, HIV-2 ST, HIV-2 SBL/ISY, BIV 127, FIV PPR, EIAV uk, and VMV 8.5 were expressed as fusions with the GAL4 BD and tested for their ability to interact with PRA1 in a mammalian two-hybrid assay. An additional transfection with vectors expressing the GAL4 BD and VP16 AD without fusion partners was included as a negative control for background levels of CAT production (BD). Error bars indicate the standard deviations of duplicate transfections. (B) Expression of each of the lentivirus envelope cytoplasmic domains fused to the GAL4 BD. The sizes (shown in kilodaltons) of each of the expression products are consistent with the expected sizes of fusions of the different lentivirus envelope cytoplasmic domains with the GAL4 BD. Lysates from transfected 293 T cells were separated on a 10% polyacrylamide gel, transferred to a nylon membrane, and detected with a GAL4 BD-specific monoclonal antibody (Clontech).