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Fig. 7 | Retrovirology

Fig. 7

From: Moloney leukemia virus 10 (MOV10) inhibits the degradation of APOBEC3G through interference with the Vif-mediated ubiquitin–proteasome pathway

Fig. 7

The DEAG-box motif of MOV10 is required for the binding of MOV10 with ElonginC or Cullin 5. a The effect of MOV10-DEAG mutant on Vif-mediated A3G degradation. 293T cells were transfected with 0.4 μg of pcDNA3.1-Vif-HA, 0.8 μg of pcDNA3.1-A3G-HA, and 1.5 μg of pcDNA3.1-MOV10-FLAG or pcDNA3.1-MOV10-DEAG-mutant-FLAG as indicated. After 48 h, cell lysates were detected by western blotting assay with anti-HA, anti-FLAG, and anti-GAPDH antibodies. Values represent portions of A3G-HA normalized against GAPDH and compared with control. b, c Co-immunoprecipitated analysis of the interaction between MOV10-DEAG mutant and ElonginC or Cullin 5. Human 293T cells were transfected with 2 μg of pcDNA3.1-MOV10-HA or pcDNA3.1-MOV10-DEAG-HA and 6 μg of pcDNA3.1-ElonginC-FLAG or pcDNA3.1-Cullin 5-FLAG. After 24 h, MG132 were added in the transfected cells for 16 h. Then, the cells were collected for co-immunoprecipitation analysis with anti-HA agarose beads and detected by western blotting with anti-HA, anti-FLAG, and anti-GAPDH antibodies. In each transfection, empty vector pcDNA3.1 was used to equalize DNA amounts. Values in a represent percentages of A3G-HA normalized against GAPDH relative to control. The bar graphs in a represent the average expression of A3G with different treatments and relative to the A3G-only reaction control (set to 100%). Data in a represent mean ± SD from three independent experiments. Statistical significance was determined using t test: **p ≤ 0.01. All the data in a, b, and c is representative of at least three independent experiments

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