Responsible for synthesis of pseudouridine from uracil-32 in cytoplasmic transfer RNAs. Involved in riboflavin biosynthesis. Converts 2,5-diamino-6-(ribosylamino)-4(3H)-pyrimidinone 5'-phosphate into 5-amino-6-(ribosylamino)-2,4(1H,3H)-pyrimidinedione 5'-phosphate. uridine(32) in tRNA = pseudouridine(32) in tRNA 2,5-diamino-6-hydroxy-4-(5-phosphoribosylamino)-pyrimidine + H(+) + H2O = 5-amino-6-(5-phospho-D-ribosylamino)uracil + NH4(+) Binds 1 zinc ion. Cofactor biosynthesis; riboflavin biosynthesis; 5-amino-6-(D-ribitylamino)uracil from GTP: step 2/4. Present with 1540 molecules/cell in log phase SD medium. In the N-terminal section; belongs to the pseudouridine synthase RluA family. In the C-terminal section; belongs to the cytidine and deoxycytidylate deaminase family.