Selection of aminoacyl-tRNAs at sense codons: the size of the tRNA variable loop determines whether the immediate 3′ nucleotide to the codon has a context effect

摘要

Codon context can affect translational efficiency by several molecular mechanisms. The base stacking interactions between a codon-anticodon complex and the neighboring nucleotide immediately 3′ can facilitate translation by amber suppressors and the tRNA structure is also known to modulate the sensitivity to context. In this study the relative rates of aminoacyl tRNA selection were measured at four sense codons (UGG, CUC, UUC and UCA), in all four 3′ nucleotide contexts, through direct competition with a programed frameshift at a site derived from the release factor 2 gene. Two codons (UGG and UUC) are read by tRNAs with small variable regions and their rates of aminoacyl tRNA selection correlated with the potential base stacking strength of the 3′ neighboring nucleotide. The other two codons (CUC and UCA) are read by tRNAs with large variable regions and the rate of selection of the aminoacyl-tRNAs in these cases varied little among the four contexts. Re-examination of published data on amber suppression also revealed an inverse correlation between context sensitivity and the size of the variable region. Collectively the data suggest that a large variable loop in a tRNA decreases the influence of the 3′ context on tRNA selection, probably by strengthening tRNA-ribosomal interactions.