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. Author manuscript; available in PMC: 2012 Mar 16.
Published in final edited form as: Mol Cell. 2011 Sep 16;43(6):962–972. doi: 10.1016/j.molcel.2011.08.008

Figure 5. Ded1 interacts directly with eIF4G1 via its third RNA binding motif.

Figure 5

A. Schematic of eIF4G1 and its protein binding motifs. B. Purified GST or full-length GST-eIF4G1 (in absence of eIF4E) in E. coli cell lysate was bound to gutathione resin prior to the addition of control buffer or 1 uM Ded1p. RNases were included in each reaction. Samples were resolved by PAGE and stained with coomassie. The input lanes showed 11.7% of the reactions in pull-down lanes. C. 1 uM of GST-eIF4G1 fragment was incubated overnight with 1 uM Ded1p and RNases before addition of glutathione sepharose. 11.1% of total input was shown for each reaction. Recombinant Ded1p was analyzed by western blot (bottom). eIF4G1 fragments were analyzed by coomassie (top); nearby ladder size in kDa is marked. The asterisk marks recombinant Ded1. See also Figure S6.