plasmid containing recombinant full-length eif4g1  (Addgene inc)

Journal: Nucleic acids research

Article Title: mRNA- and factor-driven dynamic variability controls eIF4F-cap recognition for translation initiation.

doi: 10.1093/nar/gkac631

Figure Lengend Snippet: Figure 2. eIF4G1 accelerates eIF4E–mRNA binding in an mRNA-dependent manner and allows the interaction to persist on the translation initiation timescale. (A) eIF4E-mRNA association rates in the absence (grey) and presence (red) of full-length yeast eIF4G1. (B) Fold-stimulation of eIF4E–mRNA association rate by eIF4G1, as a function of mRNA length. (C) Kinetics of eIF4E–mRNA dissociation for transient binding events in the presence (blue) and absence (grey) of eIF4G1. (D) Representative single-molecule fluorescence trace for eIF4E–mRNA binding in the presence of eIF4G1. The inset shows representative transient and prolonged events on an expanded time axis. (E) Cumulative probability distributions of eIF4E–NCE102 mRNA event durations in the absence (grey) and presence (blue) of eIF4G1, showing appearance of slowly-dissociating events when eIF4G1 is present. (F) eIF4E– mRNA dissociation rates for long-lived binding events in the presence of eIF4G1. (G) Apparent equilibrium dissociation constants for the eIF4E–mRNA interaction in the presence of eIF4G1. (H) eIF4E–mRNA association rates in the presence (red) and absence (grey) of eIF4G11–452.

Article Snippet: The plasmid containing recombinant full-length eIF4G1 was a gift from Sarah Walker (also available from Addgene as plasmid #122248).

Techniques: Binding Assay, Fluorescence