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. 2010 Jan 6;285(12):9018–9029. doi: 10.1074/jbc.M109.065698

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© 2010 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 6. — Gel shift assay and inhibition experiments with tRNA-dG18 and -dG18dG19A20 variants. A, steady-state binding of TrmH to wild-type tRNA^Phe (upper panel), tRNA-dG18 (middle panel), and tRNA-dG18dG19A20 (lower panel) variants. Transcripts incubated with increasing concentrations of TrmH protein in a binding reaction were then resolved by 6% native PAGE. Gels were stained sequentially with Coomassie Brilliant Blue and methylene blue for visualization of protein and RNA, respectively. B, inhibition of methyl transfer to wild-type tRNA in the presence of methylated tRNAs containing Gm18 (upper panel), tRNA-dG18 (middle panel), or tRNA-dG18dG19A20 (lower panel). The concentration of TrmH was fixed at 80 nm, whereas the concentrations of the tRNA variant heading each graph are 0 nm (open circles), 25 nm (closed circles), and 50 nm (open squares).