FIG 6.

MNK1 stimulation of viral translation does not require MNK1 binding to eIF4G1. (A) Dox-inducible endogenous eIF4G1 knockdown with simultaneous mock reconstitution (pcDNA5) or reconstitution with wt myc-eIF4G1-Flag or myc-eIF4G1(ΔMNK)-Flag. (B) Flag IP of lysates from the three cell lines after Dox induction (96 h). MNK1 co-IP occurred only with wt eIF4G1 reconstituted cells. (C) MNK1/mock depletion, TPA/mock stimulation, and PVSRIPO infection of the three cell lines as shown in Fig. 4A. Prior to infection, all cells were Dox induced (96 h), followed by siCtrl/siMNK1 treatment and TPA/mock stimulation, as shown. Note deficient eIF4E(S209) phosphorylation in mock-/eIF4G(ΔMNK)-reconstituted cells. Viral protein 2C levels were quantitated (4 h p.i.), and the values represent the average of 3 experiments normalized using the corresponding siCtrl-mock lanes. The error bars indicate SEM.