Fig. 7: Vif induces degradation of proximal/Vif-Bound cpzA3H molecule. | Nature Communications

Fig. 7: Vif induces degradation of proximal/Vif-Bound cpzA3H molecule.

From: HIV-1 vif mediates ubiquitination of the proximal protomer in the APOBEC3H dimer to induce degradation

Fig. 7

HEK293T cells were co-transfected with either an empty vector (no Vif) or HIV-1 Vif expression plasmids, alongside the untagged cpzA3H (WT) and myc-cpzA3H (WT or W90A) expression plasmids. Independent vectors were used for each cpzA3H construct. At 40 h post-transfection, the intracellular levels of cpzA3H were analyzed by western blotting using an anti-A3H rabbit polyclonal antibody. Vif expression levels were detected with an anti-Vif mAbs. The ß-Tub was used as a loading control. We tested NL4-3 WT Vif (lanes 5 and 6), the NL4-3 N48H Vif variant (lanes 7 and 8), and LAI Vif (lanes 9 and 10) for their ability to degrade cpzA3H. Both myc-tagged and untagged WT cpzA3H were similarly degraded by all three Vif proteins (lanes 5, 7, and 9). In contrast, degradation occurred only for WT cpzA3H, which binds to Vif, when WT and W90A cpzA3H were co-expressed (lanes 6, 8, and 10).

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