Figure 1

Interaction between the HIV Rev and the cellular LEDGF/p75 proteins under in vitro and in vivo conditions. (A) An ELISA assay system was used to determine the Rev(-GFP)-LEDGF/p75, Rev(-GFP)-IN and IN-LEDGF/p75 interactions. LEDGF/p75 or IN were bound to ELISA plate and the binding of Rev(-GFP) to LEDGF/p75 () or to IN () as well as the binding of LEDGF/p75 to IN () was estimated as described in Materials and Methods. For control, the binding of the GFP to LEDGF/p75 () and to IN () were studied. (B) Apparent K_d as calculated from the results presented in (A). (C) The BiFC method was used in yeast cells to demonstrate Rev-LEDGF/p75 interactions. Cells were transfected with the indicated expression vectors and, following incubation appearance of fluorescence, due to protein complementation, was visualized by fluorescent confocal microscopy. The Linker plasmid contains only the half GFP (GN or GC) and a linker as described in Materials and Methods. The scale bar represents 1 µm. All other details as described in Materials and Methods.