Virus Evolution Reveals an Exclusive Role for LEDGF/p75 in Chromosomal Tethering of HIV
Figure 6
In Vivo Interaction of WT and Mutant HIV-1 IN with LEDGF/p75 and eGFP-Δ325
HeLaP4 eGFP-Δ325 or HeLaP4 cells overexpressing eGFP-LEDGF/p75(K150A) in the cytoplasm were transfected with plasmids encoding WT or double mutant A128T/E170G mRFP-INs. As a negative control, cells were transfected with an mRFP expression plasmid. Interaction between eGFP-Δ325 or eGFP-LEDGF/p75(K150A) with WT mRFP-INs or A128T/E170G mRFP-INs was detected by measuring the simultaneous diffusion of the differentially labeled proteins using FCCS. The interaction is presented as the relative extent of cross-correlation (data are shown in box plots with SD for 30 measurements). A two-sample t-test was performed to prove that the different samples were drawn from different populations. The two samples are statistically different if p < 0.01. Since cross-correlation is concentration dependent, WT IN was compared with double mutant IN for binding to eGFP-Δ325 and for binding to eGFP-LEDGF/p75. The respective reduction in affinity is represented by the arrow, and the accompanying p-values are shown at the top of the figure.