Fig. 5.
The Ctu1–Ctu2 complex is required for thiolation of uridine-34 in tRNAs from yeast and nematode. (A) Purified tRNA from indicated strains was separated on 8 M urea containing polyacrylamide gels in the presence (Left) or absence (Right) of APM. Northern hybridization was performed with multiprimed probes corresponding to either tRNALYS or tRNAMET as indicated. (B) The KADA (K62A D63A) and the AXXA (C142A C145A) mutations were introduced in Ctu1-TAP expressed from pREP-4, and the resulting plasmids were transformed in a ctu1-deleted strain together with an empty vector as indicated. Growth of corresponding strains was tested at 32°C and 36°C (Top), and the expression level of Ctu1-TAP was tested by Western blot (Middle). (Bottom) Purified tRNA from indicated strains was analyzed by APM gel as in A.