Fig. 3

Vpr-mediated SIRT7 degradation is DCAF1- and CRL4-dependent. a DCAF1 (blue) and Vpr (magenta) binding interface. The complex structure of DCAF1 and Vpr was extracted from the crystal structure of DDB1/DCAF1/Vpr/UNG2 (PDB: 5JK7). DCAF1-interacting residues of Vpr, F69 and R62 are shown in stick representation. b Plasmids expressing DCAF1 were co-transfected into HEK293T with plasmids expressing wild-type (WT) or R62D/F69A mutant Vpr. Cell lysates were subjected to immunoprecipitation with anti-FLAG antibody. c Plasmids expressing SIRT7 were co-transfected with control plasmids or plasmids expressing WT or R62D/F69A mutant Vpr at escalating concentrations. Cell lysate was analyzed by immunoblotting. d Plasmids expressing SIRT7 or Vpr were co-transfected into HEK293T cells. 40 h later, cells were treated with 1 μM MLN4924 and incubated for an additional 8 h. Cell lysate was analyzed by immunoblotting. All experiments were repeated two to three times with similar results