Figure 4.
Cup mediates an indirect interaction between Smg and eIF4E. Labeled eIF4E as well as wild-type and mutant Cup proteins were generated via coupled transcription/translation in rabbit reticulocyte lysate in the presence of 35S-methionine. The eIF4E was mixed with either of the Cup proteins in the presence of GST-Smg583–763 and glutathione agarose. A sample of the in vitro-translated eIF4E (load) representing 30% of protein used in the captures and the elutions from the captures in the presence of wild-type Cup or CupY342A/L379A/L383A are shown.