Figure 4.
Characterization of the tRNA MTase activity of recombinant TrmK protein using radiolabeled in vitro transcripts of B. subtilis tRNASer. The transcripts were incubated in the presence of 5 μg of B. subtilis TrmK enzyme and AdoMet. The transcripts were then digested by either nuclease P1 (A) or RNase T2 (B) and the resulting nucleotides were analyzed by 2D-TLC on cellulose plates and autoradiography. The nature of the labeled triphosphate nucleoside and the enzyme used to hydrolyze the transcripts are given above the autoradiographs. Circles in dotted lines show the migration of the canonical nucleotides used as UV markers. m1A > p is for 1-methyladenosine 2′–3′ cyclic phosphate.