RNA-binding activity of translation initiation factor eIF4G1 from Saccharomyces cerevisiae

In vitro translation of GST-eIF4G1 fusion proteins in an eIF4G-dependent extract. Methionine incorporation at 25°C was measured as described previously (Dominguez et al. 1999) using 10-μL reaction mixtures containing 5 μg total yeast RNA as mRNA source and 3.6 μCi ³⁵S-methionine. The kinetics of methionine incorporation for four proteins is shown on the left. On the right, stimulation of translation is indicated by ++ (like GST-eIF4G1_1–952 and GST-eIF4G1_188–952); +, intermediate; -, like “no factor added.”