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. 2017 Mar;23(3):406–419. doi: 10.1261/rna.059667.116

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© 2017 Han et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society

This article is distributed exclusively by the RNA Society for the first 12 months after the full-issue publication date (see http://rnajournal.cshlp.org/site/misc/terms.xhtml). After 12 months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.

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FIGURE 7. — ORF240 catalyzes efficient m³C modification of tRNA^Thr(IGU) but inefficient modification of tRNA^Ser(CGA). (A) Titration of ORF240 m³C methyltransferase activity on tRNA^Thr(IGU) and tRNA^Ser(CGA). Of note, 0.35 pmol of tRNA^Thr(IGU) or tRNA^Ser(CGA) purified from a trm140Δ strain was incubated with serial fivefold dilutions of ORF240 (2.5, 0.5, 0.1, 0.02, 0.004 µM) purified from E. coli or with buffer (−) in the presence of SAM at 30°C for 1 h, and tRNA was analyzed for m³C by primer extension in the presence of ddTTP, as described in Materials and Methods. (B) Plot of m³C modification as a function of the concentration of ORF240 in A.