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. 2013 Mar 15;27(6):627–638. doi: 10.1101/gad.212738.112

Figure 3.

Figure 3.

A strong Py tract and U2AF59 favor completion of the second step of splicing. (A) TER1 mutants with strengthened (Py+) or weakened (Py−) pyrimidine tract or cytosines replaced with uridines (C2U) were integrated at the ter1 locus. (B) Detection of precursor, spliced, and cleaved forms by Northern blot. Loading control (LC) and quantification are as in Figure 2B, with n = 2. (C) Northern analysis of RNA from cells containing the ura4 reporter with TER1 intron in the context of prp2+ (U2AF59) and prp2D307N mutant, respectively. Cells were grown at 25°C to 5 × 106 cells per milliliter, shifted to 36°C, and harvested after 0 or 30 min. (Lane 1) Quantification of cleaved and spliced forms relative to LC (snR101) for each lane and normalized to wild type. Quantification was performed as in Figure 2B, with n = 2.