Figure 2. TRM140a and ADAT2/3 are required for methylation and editing of tRNAThrAGU.
a, Primer extension as in Fig. 1a with a tRNAThr transcript. The presence of absence of recombinant enzymes and SAM is indicated. ‘Primer’ refers to a primer-alone reaction. b, A radioactive version of the transcript in a incubated with ADAT2/3, TRM140a and SAM (left panel), the methyltransferase and SAM but no deaminase, or no enzyme. pC, 5′ cytosine monophosphate; Pi, inorganic phosphate. c, Reincubation of m3C-containing tRNA with TRM140a and ADAT2/3 (left panel) and ADAT2/3 (no methyltransferase) (right panel); all reactions contain SAM. The data are representative of at least three independent experiments.