Conformational-Dependent and Independent RNA Binding to the Fragile X Mental Retardation Protein
Figure 2
FMRP interacts weakly with annealed BC1 RNA. Equal amounts of 35S-FMRP was titrated with various amounts of (a) unannealed biotinylated-BC1 RNA, (b) annealed biotinylated-BC1 RNA, and (c) unannealed biotinylated-eEF-1A RNA, as indicated. Bound (B) and unbound (U) 35S-FMRP was recovered, resolved by SDS-PAGE and subject to autoradiography. Nonspecific binding to the SoftLink avidin resin is shown in the -RNA lanes. (d) Graphical analysis of the above data for 4 reactions per concentration. (e) Binding of annealed or unannealed eEF1A RNA (60 nM) to 35S-FMRP. The percent binding corrected for background of 4 reactions per RNA type is plotted ( by ANOVA).
