An Interaction between KSHV ORF57 and UIF Provides mRNA-Adaptor Redundancy in Herpesvirus Intronless mRNA Export
Figure 7
Knockdown of both Aly and UIF impairs the ability of ORF57 form an export competent viral ribonucleoprotein particle.
(A) 293, 293ΔAly, 293ΔUIF and 293ΔAlyΔUIF cells were mock treated or treated with 2 µg/ml doxycyclin grown for 72 h. Cell lysates were analysed by western blotting using Aly-, UIF- and UAP56-specific antibodies. (B) (i) 293 cells were transfected with pEGFP and grown for a further 24 h. Cell lysates were incubated with GFP-TRAP-Affinity beads and after washing, the precipitated proteins were detected by western blotting using GFP-, UAP56-, FSAP79- and TAP-specific antibodies. (ii) 293, 293ΔAly, 293ΔUIF and 293ΔAlyΔUIF cells were treated with 2 µg/ml doxycyclin and grown for 48 h before being transfected with pGFP-ORF57 and grown for a further 24 h. Cell lysates were incubated with GFP-TRAP-Affinity beads and after washing, the precipitated proteins were detected by western blotting using GFP-, UAP56-, FSAP79- and TAP-specific antibodies. Transfected cell lysates were used as an input control.