CO₂-sensitive tRNA modification associated with human mitochondrial disease

It has been generally thought that tRNA modifications are stable and static, and their frequencies are rarely regulated. N⁶-threonylcarbamoyladenosine (t⁶A) occurs at position 37 of five mitochondrial (mt-)tRNA species. We show that YRDC and OSGEPL1 are responsible for t⁶A37 formation, utilizing L-threonine, ATP, and CO₂/bicarbonate as substrates. OSGEPL1-knockout cells exhibit respiratory defects and reduced mitochondrial translation. We find low level of t⁶A37 in mutant mt-tRNA isolated from the MERRF-like patient's cells, indicating that lack of t⁶A37 results in pathological consequences. Kinetic measurements of t⁶A37 formation reveal that the Km value of CO₂/bicarbonate is extremely high (31 mM), suggesting that CO₂/bicarbonate is a rate-limiting factor for t⁶A37 formation. Consistent with this, we observe a low frequency of t⁶A37 in mt-tRNAs isolated from human cells cultured without bicarbonate. These findings indicate that t⁶A37 is regulated by sensing intracellular CO₂/bicarbonate concentration, implying that mitochondrial translation is modulated in a codon-specific manner under physiological conditions.