Figure 2
From: High-affinity consensus binding of target RNAs by the STAR/GSG proteins GLD-1, STAR-2 and Quaking
STAR-2 binding to TGE RNA and determination of STAR-2 binding specificity. A. Wild-type TGE RNA compared to sequences for C21A TGE and TGE with changes in the upstream 5'-UAA element. B. Electrophoretic gel mobility shift titration of the STAR-2 STAR Domain into radiolabeled TGE RNA with STAR-2 concentration and bound complex denoted. The equilibrium dissociation constant and Hill values shown were calculated from the plot of fraction bound as a function of STAR-2 concentration. C. STAR-2 STAR Domain binding to fluorescein-labeled TGE RNA measured by fluorescence-polarization. Kd and Hill values shown were determined from the fit of the polarization versus concentration of STAR-2. Error bars represent the standard deviation of 10 consecutive readings from the same plate. D. Plot of the normalized polarization against concentration of mutant 12-mer competitor RNAs. Traces display the competitive efficiency of three point mutants, wild-type TGE half-site and self-competition with the 28-nt TGE. E. Table of relative IC50 values for the mutant 12-mer RNA sequences compared to wild-type TGE half-site.