Formation of pseudouridine at positions 27 and 28 in the anticodon stem and loop of transfer RNAs; at positions 34 and 36 of intron-containing precursor tRNA(Ile) and at position 35 in the intron-containing tRNA(Tyr) (PubMed:9671058, PubMed:10356324, PubMed:25219674). Catalyzes pseudouridylation at position 44 in U2 snRNA (PubMed:25219674). Also catalyzes pseudouridylation of mRNAs (PubMed:25219674, PubMed:31477916). a uridine in tRNA = a pseudouridine in tRNA uridine in snRNA = pseudouridine in snRNA a uridine in mRNA = a pseudouridine in mRNA Binds 1 zinc ion per subunit. kcat is 0.4 min(-1) for tRNA(Val) (PubMed:10356324). kcat is 0.5 min(-1) for tRNA(Ile) (PubMed:10356324). Present with 8130 molecules/cell in log phase SD medium. Belongs to the tRNA pseudouridine synthase TruA family.