Mutation of RNA-contacting amino acid residues adversely affect PUS1 activity.

(a) Table showing the average percent of uridine-to-Ψ conversion of the reactions shown in Fig 3C (from two independent replicates), and the percentage of PUS1 activity, normalized by the percent pseudouridylation in the PUS1_wt reaction. (b) Protein gel comparing the input amount and purity of the PUS1 variants in the in vitro reactions. (c) Electrophoretic mobility shift assays of the mutant PUS1 enzymes (PUS1_R132A left, PUS1_R362A right).

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