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Figure 3
Mutation of the U6 ISL does not substantially affect binding between U6 and Prp24. (a) An electrophoretic mobility shift assay (EMSA) was used to assess the binding affinity of Prp24 towards U6 snRNA nucleotides 30–101 with a U100C/U101C mutation or the same RNA with an additional A62G mutation. The total concentration of labeled U6 is 0.5 nM in all samples. (b) Plot of bound RNA as a function of protein concentration. The A62G mutant exhibits a higher fraction of bound RNA, suggesting that stabilization of the ISL increases the fraction of U6 RNA that is competent to bind Prp24. Data points denoted by red arrows are likely to correspond to weak binding of misfolded U6 and were not used for calculation of the binding constants shown here. All data are derived from four independent technical replicates, except for those at 2400 nM, which were performed in duplicate.

Journal logoSTRUCTURAL
BIOLOGY
ISSN: 2059-7983
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