Discovery and characterization of tRNA Ile lysidine synthetase (TilS)

In the bacterial decoding system, the AUA codon is deciphered as isoleucine by tRNA Ile bearing lysidine (L, 2‐lysyl‐cytidine) at the wobble position. Lysidine is an essential modification that determines both the codon and amino acid specificities of tRNA Ile . We identified an enzyme named tRNA Ile lysidine synthetase (TilS) that catalyzes lysidine formation by using lysine and ATP as substrates. Biochemical studies revealed a molecular mechanism of lysidine formation that consists of two consecutive reactions involving the adenylated tRNA intermediate. In addition, we deciphered how Escherichia coli TilS specifically discriminates between tRNA Ile and the structurally similar tRNA Met , which bears the same anticodon loop. Recent structural studies unveiled tRNA recognition by TilS, and a molecular basis of lysidine formation at atomic resolution.