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. Author manuscript; available in PMC: 2023 Feb 3.
Published in final edited form as: Mol Cell. 2022 Jan 19;82(3):645–659.e9. doi: 10.1016/j.molcel.2021.12.023

Figure 4. Multiple PUS pseudouridylate pre-mRNA sequences.

Figure 4.

a) Schematic of in vitro pseudouridylation assay with RNA made from a pool of 6000 oligos containing all the sites identified in HepG2 chromatin-associated RNA. In vitro pseudouridylation was carried by incubating pool RNA with recombinant human pseudouridine synthases (PUS) and pseudouridines were identified by Pseudo-seq. b) Genome browser view of Pseudo-seq reads at pseudouridine sites following RNA incubation with a recombinant PUS or no PUS control. Plots for three intronic pre-mRNA pseudouridines: a PUS1 target GPC3, PUS7 target in RBM39 and a TRUB1 target in NOMO2. c) Combined distribution of pseudouridines validated as direct targets of all tested PUS by in vitro Pseduo-seq assay. d) Summary of pseudouridines assigned as direct targets of each PUS protein from in vitro Pseudo-seq assay. e) Weblogo summarizing frequency of motifs identified among targets of PUS7 and TRUB1.