biostudies-literatureUnknown18(12)Li HMethylation is a common post-transcriptional modification of tRNAs, particularly in the anticodon loop region. The cytosine 38 (C38) in tRNAs, such as tRNA^Asp-GUC, tRNA^Gly-GCC, tRNA^Val-AAC, and tRNA^Glu-CUC, can be methylated by human DNMT2/TRDMT1 and some homologs found in bacteria, plants, and animals. However, the substrate properties and recognition mechanism of DNMT2/TRDMT1 remain to be explored. Here, taking into consideration common features of the four known substrate tRNAs, we investigated methylation activities of DNMT2/TRDMT1 on the tRNA^Gly-GCC truncation and point mutants, and conformational changes of mutants. The results demonstrated that human DNMT2/TRDMT1 preferred substrate tRNA^Gly-GCC in vitro. L-shaped confRNA biology2531-2545https://www.ebi.ac.uk/biostudies/studies/S-EPMC8632113biostudies-literatureNew substrates and determinants for tRNA recognition of RNA methyltransferase DNMT2/TRDMT1.PMC8632113Li HWu JCai CQin MMa YChen YZhu DDai HfalseNew substrates and determinants for tRNA recognition of RNA methyltransferase DNMT2/TRDMT1.Methylation is a common post-transcriptional modification of tRNAs, particularly in the anticodon loop region. The cytosine 38 (C38) in tRNAs, such as tRNA^Asp-GUC, tRNA^Gly-GCC, tRNA^Val-AAC, and tRNA^Glu-CUC, can be methylated by human DNMT2/TRDMT1 and some homologs found in bacteria, plants, and animals. However, the substrate properties and recognition mechanism of DNMT2/TRDMT1 remain to be explored. Here, taking into consideration common features of the four known substrate tRNAs, we investigated methylation activities of DNMT2/TRDMT1 on the tRNA^Gly-GCC truncation and point mutants, and conformational changes of mutants. The results demonstrated that human DNMT2/TRDMT1 preferred substrate tRNA^Gly-GCC in vitro. L-shaped conf2021-01-01T00:00:00Z2021 Dec2026-05-06T03:13:39.34Z2025-02-19T00:56:14.191ZS-EPMC86321133411097510.1080/15476286.2021.1930756