Background: tRNA methyltransferases specifically recognize substrate tRNAs. Results: To clarify the tRNA recognition mechanism of TrmI, three tRNA species and 45 variants were analyzed in vitro and in vivo. Conclusion: TrmI recognizes the aminoacyl stem, variable region, C56, purine 57, A58, and U60 in the T-loop of tRNA. Significance: Our in vitro experimental results explain the regulation of in vivo methylation levels in tRNAs.
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