HPLC chromatograms of tRNA hydrolysates from the wild-type (A) and iscS51 mutant (B) strains. The nucleosides were monitored at 295 nm to maximize the detection of all thiolated nucleosides. mnm5s2U, cmnm5s2U, s2C, ms2io6A, s4U, ms2i6A, pseudouridine (Ψ), and N-6-isopentenyladenosine (i6A) were identified by comparing UV spectra with published spectra (13). For s2C, the molecular weight of the protonated form was determined by mass spectrometry (Jäger et al., unpublished data). AU, absorbance units.