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. Author manuscript; available in PMC: 2019 Aug 15.
Published in final edited form as: Science. 2019 Jan 31;363(6428):710–714. doi: 10.1126/science.aaw5569

Figure 3. FAM32A promotes exon ligation by binding the 5’-exon.

Figure 3

FAM32A promotes exon ligation by binding the 5´-exon. (A and B) Depletion of FAM32A impairs exon ligation. (C) Overview of the UV cross-linking experiment. Note that C(-2) was changed to U(-2) for these experiments. (D) FAM32A promotes exon ligation. (E) Effect of FAM32A depletion on exon ligation efficiency. Experiments were performed using a substrate with a single 32P at U(-2) of the 5´-exon. Error bars represent standard deviation (n=3). (F) C* complexes accumulate in FAM32A-depleted extracts. Shown is RNA extracted from affinity-purified P complexes (see also fig. S8). (G) FAM32A cross-links to the 5´-exon. SDS-PAGE of proteins labeled through cross-linking. SII-FAM32A, Strep-tactin-tagged FAM32A; 32p, 32P radioactive phosphate; 4SU, 4-thio-uridine. (H) Positioning of FAM32A and Prp8 around C(-2) of the 5´-exon rationalizes the cross-linking results.