Fluorimetric distance determination by resonance energy transfer. Ribosome-bound transfer RNA

Abstract

Using the technique of singlet-singlet (Förster-type) resonance energy transfer, we have determined five distances in the programmed ribosome, either with the P site or with both the A and the P sites occupied. Two of the distances are new and two agree with earlier measurements; the fifth showed disagreement in detail with earlier results of others, but a consistent general trend. The distances substantiate a current model for the location of ribosomally bound tRNA, except in regard of the position of the 3' end of P-site tRNA, which seems according to our results to lie too far away from the 3' terminus of the 16 S RNA to be accommodated in the model. We present new evidence for the hypothesis that anomalously charged tRNA does not bind to the cognately programmed A site in the same way as does tRNA charged with an amino acid. Occupation of the A site restricts mobility of the 3' end of tRNA in the P site.