A Structural Motif for mRNA Target Recognition by a tRNA Pseudouridine Synthase

We used an in vitro pool based strategy to probe the sequence and structural requirements for mRNA target recognition by the yeast tRNA pseudouridine synthase Pus1. Overall design: RNA in vitro transcribed from synthetic oligo pools was pseudouridylated with a source of Pus1 activity. Pseudouridylation was measured using Pseudo-seq, a seqeuncing based technique for identifying ?s (Carlile et al. Nature 2014).