FIGURE 2.
Both MUS81 and its cofactors EME1 and EME2 are down-regulated by Vpr from multiple species. A and B, MUS81 and EME1 plasmids (A) or MUS81 and EME2 plasmids (B) were transiently co-transfected with increasing amounts of VprHIV-1 NL4-3 plasmid. At 48 h post-transfection, cells were harvested and analyzed by Western blotting. C, endogenous levels of MUS81, EME1, and EME2 were analyzed with increasing amounts of VprHIV-1 NL4-3. D and E, MUS81 and EME1 plasmids were co-transfected with increasing amounts of plasmid encoding Vpr isolated from SIV that infects chimpanzee, Vprcpz-PTT and Vprcpz-PTS (D), or encoding Vpr from HIV-1 group M, N, or O, VprHIV-1 M, VprHIV-1 N, and VprHIV-1 O (E). All experiments were repeated three times with equivalent results. F, sequence alignment of the Vpr proteins used in A–D was performed with Clustal Omega (68) and colored by ESPript 3.0 (69).