Figure 2. Effects on Brr2-mediated U4/U6 unwinding and interplay with other U4/U6 proteins.

(A) Native gels monitoring yU4/U6 di-snRNA unwinding by yBrr2 in the absence of other proteins (top), in the presence of yPrp3^CTF (middle) or yPrp3^DUF1115 (bottom). Asterisks indicate radioactive label on yU4 snRNA. (B) Quantification of the data in (A). The data were fit to a single exponential equation: % duplex unwound = A{1 − exp(−k_u t)}; A—amplitude of the reaction; k_u—apparent first-order rate constant; t—time. Amplitudes and rate constants are listed. Errors indicate standard errors of the mean of four independent experiments. (C) Binding of yPrp3^DUF1115 (left three lanes of each panel) or yPrp3^CTF (right three lanes of each panel) to yU4/U6^fused (lanes 1–6; left), yU4/U6^fused pre-bound to Snu13 (lanes 7–12; middle) or yU4/U6^fused pre-bound to Snu13 and Prp31¹⁻⁴⁶² (lanes 13–18; right) under otherwise identical conditions. All three panels are from the same gel and were regrouped. Schemes of yU4/U6^fused alone or pre-bound by proteins are shown on the top.

DOI: http://dx.doi.org/10.7554/eLife.07320.006