tRNA-tRNA interactions within cellular ribosomes.

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Abstract
We describe an assay that converts the effects of tRNA-tRNA contacts at two particular codons into a quantitative effect on .beta.-galactosidase level. The assay measures the separate and combined efficiency of suppression at adjacent nonsense codons in vivo using a set of specially created homologous messages. In a survey of distal anticodon arm substitutions, we find that particular mutant tRNAs occupying the P-site reduce the apparent efficiency of the suppressor tRNA reading the A-site codon by factors of 2-170. By using measured tRNA-tRNA distances and the crystallographic tRNA structure, we proposed a model of the tRNA-tRNA-mRNA complex. In the model, the anticodon loops of the P-site and A-site tRNAs contact one another in a way that is consistent with our combined tRNA efficiency data. These results suggest that tRNA tRNA interactions that modulate tRNA action are an inevitable feature of translation.