Dnmt2 is a methyltransferase with high sequence similarity to various enzymes from the eukaryotic DNA:methyltransferase family, which includes in particular Dnmt1 and Dnmt3 enzymes. Although it uses the same catalytic residues as other Dnmt enzymes, DNA methylation activity producing dm5C by Dnmt2 is weak, and both its existence and biological role are controversial. Instead, the enzmye displays robust tRNA Methylation activity to yield m5C at position 38 of tRNAs in vitro and in vivo. Mammalian substrates include tRNAs Asp, Gly, and Val. Dnmt2 homologues are widespread in eukaryotes, including certain single cellular eukaroytes, but only one bacterial homologue is known. Active enzymes have been confirmed in mammalia, Drosophila melanogaster, Saccharomyces pombe, and Entamoeba histolytica.