Chemical Accessibility of 18S rRNA in Native Ribosomal Complexes: Interaction Sites of mRNA, tRNA and Translation Factors

Abstract

During protein synthesis the ribosome interacts with ligands such as mRNA, tRNA and translation factors. We have studied the effect of ribosomeligand interaction on the accessibility of 18S rRNA for single strandspecific modification in ribosomal complexes that have been assembled in vivo, i. e. native polysomes. A comparison of the modification patterns derived from programmed and nonprogrammed ribosomes showed that bases in the 630- and 1060- loops (530- and 790-loops in E. coli) together with two nucleotides in helices 33 and 34 were protected from chemical modification. The majority of the protected sites were homologous to sites previously suggested to be involved in mRNA and/or tRNA binding in prokaryotes and eukaryotes, implying that the interaction sites for these ligands are similar, if not identical, in naturally occurring programmed ribosomes and in in vitro assembled ribosomal complexes. Additional differences between programmed and nonprogrammed ribosomes were found in hairpin 8. The bases in helix 8 showed increased exposure to chemical modification in the programmed ribosomes. In addition, structural differences in helices 36 and 37 were observed between native 80S runoff ribosomes and 80S ribosomes assembled from isolated 40S and 60S subunits.