FIGURE 1.
Pseudouridine sequencing of mitochondrial tRNAGlu. A, schematic of pseudouridine sequencing shown in the cloverleaf structures of the human mitochondrial tRNAGlu (WT and mutant (MT). Solid lines represent the DIG-labeled oligonucleotide probe specific for tRNAGlu. Broken lines represent the potential stops of reverse transcription reaction caused by base modification, such as CMC-pseudouridine and m1A. B, 2 μg of mitochondrial RNA from control (C3 and C4) and mutant (III-6 and III-9) cells was incubated with CMCT for CMC modification of Ψ residues. Reverse transcription was carried out to identify the stops caused by CMC-pseudouridine. The tRNAGlu transcript was used as a control without base modification. Marker, DIG-labeled oligonucleotides of variable length.