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. 2017 Mar;23(3):406–419. doi: 10.1261/rna.059667.116

FIGURE 4.

FIGURE 4.

An XGU anticodon and t6A37 are sufficient for ORF240 binding and for m3C modification of tRNAPhe. (A) Schematic of tRNAPhe(GAA)29,50 variant. The secondary structure of tRNAPhe is shown, with highlighting of base pairs 29:41 and 50:64 that were flipped, and bold outline of residues N34–37 that were changed from GAAG to CGUA. (B) A CGU anticodon and t6A37 are sufficient for binding of tRNAPhe to ORF240. Strains with integrated tF(GAA)29,50 variants as indicated were analyzed by the ORF240 tRNA pull-down assay as described in Figure 3C. (C) Summary of pull-down results in B. (D) A CGU anticodon and t6A37 are sufficient for m3C modification of tRNAPhe. Bulk RNA from strains without (lane 1) or with a tRNAPhe(GAA)29,50 [CGUA]34-37 variant (lane 2) was analyzed by primer extension for m3C, using a primer complementary to residues 52–36 of the variant. Lane 3, primer alone.