It was unknown whether a synthetic Shine-Dalgarno (SD) oligonucleotide labelled with <$E nothing sup 32 roman P> at its 5'-end ([<$E nothing sup 32 roman P>]oct) would be able to reach the anti-SD sequence of l6S rRNA at the early stages of translation only or during elongation. To verify this, [<$E nothing sup 32 roman P>]oct was incubated with 30S ribosomal subunits (RSUs), 70S ribosomes and polysomes, separately, while the SD/anti-SD binding was checked in them through sucrose gradients. The anti-SD sequence resulted highly available in 30S RSUs and sufficiently available in ribosomes. In both 30S RSUs and ribosomes, the addition of a model 002 mRNA in equimolar proportions displaced [<$E nothing sup 32 roman P>]oct for about 50 %. However, in ribosomes the presence of initiation factors (IFs) and fMet-tRNA influence neither the binding of [<$E nothing sup 32 roman P>]oct nor the competition coming from mRNA. In polysomes, [<$E nothing sup 32 roman P>]oct was unable to hybridize the anti-SD sequence, in agreement with the hypothesis that mRNA and 16S rRNA are involved in the SD/anti-SD interaction also during elongation.

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