The RING-CH Ligase K5 Antagonizes Restriction of KSHV and HIV-1 Particle Release by Mediating Ubiquitin-Dependent Endosomal Degradation of Tetherin
Figure 8
Tetherin cytoplasmic tail lysine residues are ubiquitinated in the presence of K5 and Vpu.
HeLa cells were transiently transfected with the indicated THN-HA expression vector in combination with K5, Vpu or GFP and in the presence or absence of 6His-tagged ubiquitin. 48h after transfection, cells were treated for 8 hours with BafA1 (100nM) to prevent tetherin degradation. Cell lysates were then harvested and ubiquitinated proteins were isolated by binding to Ni-Nti-agarose. Cell lysates and pull-downs were analyzed by western blot for THN-HA detecting the HA tag.
doi: https://doi.org/10.1371/journal.ppat.1000843.g008