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. 2021 Apr;27(4):420–432. doi: 10.1261/rna.077198.120

FIGURE 3.

FIGURE 3.

Disease-linked mutations in human (mt)pre-tRNALeu(UUR) decrease methylation efficiency catalyzed by the human mtRNase P complex. (A) Single-turnover methylation rates of mutant (mt)pre-tRNAIle variants relative to that of the wild-type (mt)pre-tRNAIle catalyzed by the MRPP1/2 subcomplex using standard reaction conditions. Reaction contained 800 nM unlabeled pre-tRNA and 5 µM MRPP1/2. (B) Quantification of methylation efficiency of mutant (mt)pre-tRNALeu(UUR) variants relative to wild-type after 60 min incubation with mtRNase P. Reaction contained 800 nM unlabeled pre-tRNA and 5 µM MRPP1/2 and MRPP3. (C) Representative gel showing methylation efficiencies of wild-type and mutant (mt)pre-tRNALeu(UUR) variants by human mtRNase P at 30 and 60 min using standard reaction conditions. Black arrows indicate two products of primer extension reactions; the read through (unmethylated pre-tRNA) and the “run-off” at the methylation sites (methylated pre-tRNA).