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. 2021 Dec 6;49(22):13045–13061. doi: 10.1093/nar/gkab1148

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© The Author(s) 2021. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (https://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Figure 8. — G/I34 is the prerequisite for m⁵C38 formation by mammalian Dnmt2s. The capacity of orthologs of tRNA^Gly(GCC) from different species and their -G34A mutants to be methylated by their cognate Dnmt2 from M. musculus (A), D. rerio (B) and D. melanogaster (C), respectively. (D) Summary of the known tRNA substrates of Dnmt2s from different species. Our results, together with those of previous studies, showed that G/I34 in tRNA substrates is required for mammalian Dnmt2 recognition; while in other lower eukaryotes and prokaryotes, G/I34 is dispensable for Dnmt2 recognition. The tRNAs labeled with * can only be methylated by the cognate Dnmt2 in vitro, but not in vivo.