Fig. 4

SIRT7 degradation is a conserved feature of HIV-1 Vpr. a Plasmids expressing SIRT7 were co-transfected with increasing amounts of plasmids expressing HIV-1 NL4-3 Vpr or SIVmac Vpx. Cell lysate was prepared and subjected to SDS-PAGE and immunoblotting. b In vitro ubiquitination assays of SIRT7 with CRL4-DCAF1c, CRL4-DCAF1c-Vpr (HIV-1 NL4-3), or CRL4-DCAF1c-Vpx (SIVmac). Poly-ubiquitinated SIRT7 species are indicated (SIRT7-Ubn). c–e Plasmids expressing SIRT7 were co-transfected into HEK293T cells with increasing amounts of plasmids expressing HIV-1 YU2, LAI, Q23, M, N, or O Vpr (c), SIVcpz PTT or PTS Vpr (d), or HIV-1 NL4-3, HIV-2 A1, HIV-2 A2, or HIV-2 B Vpr (e). Relative intensity of bands corresponding to SIRT7 was quantified (e). f, In vitro ubiquitination assays of SIRT7 with CRL4-DCAF1c in complex with HIV-1 LAI, YU2, M, or SIVcpz PTT Vpr. g In vitro ubiquitination assays of SIRT7 with CRL4-DCAF1c or CRL4-DCAF1c in complex with HIV-1 NL4-3 Vpr, SIVmac Vpx or HIV-2 A1 Vpr. All experiments were repeated two to three times with similar results