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. Author manuscript; available in PMC: 2021 Apr 28.
Published in final edited form as: Nature. 2020 Oct 28;587(7835):663–667. doi: 10.1038/s41586-020-2854-z

Fig. 3. MTR1-catalysed methylation of tRNA.

Fig. 3

a. Native m1A sites at positions 9, 22 and 58 are shown in a generic tRNA scaffold. b. In vitro transcribed tRNAs were incubated with the corresponding complementary ribozymes in presence (+) or absence (−) of m6G. The installation of m1A was probed by primer extension experiments. Primer binding sites are indicated on the tRNA scheme. Sequencing reactions were run in parallel to assign the position of the abort bands. c. Total E.coli tRNA was incubated with a tRNAAsp-A58-specific MTR1, and specific methylation of tRNAAsp was probed by primer extension with six tRNA-specific primers P4-P9. Sequences of tRNAs are given in Extended Data Fig. 6. d. LC-MS analysis of MTR1-catalyzed methylation of total E.coli tRNA. Extracted ion chromatograms (EIC, detecting MH+ (m/z 282.11±0.05), corresponding to methylated adenosines) are shown for digested tRNAs (before and after treatment with MTR1), and for synthetic m1A and m6A nucleosides.