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. 2023 Dec 5;52(3):1374–1386. doi: 10.1093/nar/gkad1136

Figure 4.

Figure 4.

The effect of tRNA modifications on decoding by E. coli tRNAGly UCC. (A) Schematic depiction of splinted ligation of differently modified chemically synthesized RNA oligonucleotides to generate full-length site-specifically modified tRNAs. (B) Decoding efficiency of the four synonymous glycine codons within a small mRNA by partly or fully modified E. coli tRNAGly UCC. [35S]-Met labelled peptides were quantified and normalized to the peptide yields resulting from the GGA containing mRNA. (C) Translation efficiency of tRNAs harboring different modifications depicted in (D) at position 34. Small mRNAs either containing GGA or GGG codons were in vitro translated in presence of [35S]-Met and the respective modified E. coli tRNAGly UCC. Peptide yields were normalized to the sample of unmodified tRNAGly UCC. Open circles indicate datapoints shown in the representative gels. (B, C) Mean of individual datapoints with standard deviation is shown. Significance was tested by ANOVA ((B) 0.00158 (left); 0.00677 (right); (C) not significant (left); 0.0204 (right) followed by Dunnett's test (0.001 ‘***’ 0.01 ‘**’ 0.05 ‘*’ >0.05 ‘ns’).