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. 2019 Dec 24;5:63. doi: 10.1038/s41421-019-0136-4

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — a Olignonuceotides (14-mer) used as substrates. b MettL3-14 is not active on dsDNA. The enzyme concentration [E] is noted in each panel. c, d MettL3-14 is active on ssDNA by two independent assays: incorporation of tritium from ³H-SAM into DNA substrate (panel c) and formation of byproduct SAH in a bioluminescence assay (panel d). Inserted box listed ssDNA sequence alignment. e Substitution of conserved cytosine within GGACT diminished Mettl3-14 activity. f Replacement of the second guanine-to-adenine (GAACT, red) or the last thymine-to-cytosine (GGACC, blue) retained comparable activity. The two lines fell on top to each other, giving rise to purple. g Comparison of CcrM (GAnTC) and MettL3-14 (GGACT) on ssDNA oligo 6 T and its derivatives. h Comparison of MettL3-14 on oligo 6 T and its corresponding RNA. i Comparison of the kinetic parameters on ssDNA and ssRNA, derived from Supplementary Fig. S2d. j Quantitative measurement of N6mA by mass spectrometry, derived from Supplementary Fig. 3. k, l Comparison of CcrM and MettL3-14 on oligo 6 T. m, n MettL3-14 is active on 28-bp dsDNA-containing mismatched pairs. Data represent the mean ± SD of three independent determinations (N = 3) performed in duplicate.