Figure 1
From: The four Zn fingers of MBNL1 provide a flexible platform for recognition of its RNA binding elements
The importance of GC dinucleotides in the context of single-strand RNA for MBNL1 binding. A and E) Sequence of RNAs used in the gel shift assays to determine the specificity for the GC dinucleotides. GC dinucleotides are shown in bold with the subsequent mutations underlined. B, C and F) Representative gel shift assays. RNAs bound to MBNL1 (2-260) are labeled on the left. Concentrations of either MBNL1 or competitive RNA in each lane are at the top of the columns. D) Representative competition assay. Each lane contains 33.3 nM MBNL1, 0.02 nM radioactive 2GC RNA and increasing concentrations of competitive RNA. The identity of the competitive RNA is labeled on the left side of the gels and concentration of the competitive RNA (nM) is shown at the top of the column. On the left of each gel is the Kd based upon a competition model of binding. G) Thermal denaturation analysis from representative RNAs monitored at 260 nm. The black line is 1GC#2, red is 2GC#1, blue is 3GC#3, green is 4GC, orange is (CUG)4, and purple is 2GC. The (CUG)4 sequence is GCUGCUGUUCGCUGCUG. Refer to part A and E for the sequences of the other RNAs.