Adjacent codon‐anticodon interactions of both tRNAs present at the ribosomal A and P or P and E sites

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Abstract
A labeled tRNA present at the A, P or E site can be partially chased from the ribosome, a cognate non‐labeled tRNA as chasing substrate being 3‐12‐times more efficient than non‐cognate tRNA at a molar ratio tRNA: 70 S = 10:1. These findings indicate that a tRNA bound to a programmed ribosome undergoes codon‐anticodon interaction at all three sites (A, P and E site). Furthermore, both labeled tRNAs present on the ribosome can be chased more effectively with cognate than with non‐cognate substrate at the same time. This finding provides strong evidence that both tRNAs present on the ribosome exhibit simultaneous codon‐anticodon interaction. This is valid for both the pretranslocational state (Ac[3H]Lys‐tRNALys in the A and [14C]tRNALys in the P site) as well as the posttranslocational state (Ac[3H]Lys‐tRNALys in the P and [14C]tRnaLys in the E site).