Mutations upstream of the ribosome-binding site affect translational efficiency.
The DNA coding for the major outer membrane lipoprotein of Escherichia coli has been fused to the coding region of the beta-galactosidase gene to measure the effect of various mutations on the efficiency of translation initiation. The various mutants were made by either inserting or deleting a small number of nucleotides into or from a region just upstream of the ribosome-binding site. These small mutations dramatically affect translation initiation as measured by the production of beta-galactosidase. We postulate that these mutations affect translation initiation by altering the secondary structure of the messenger RNA. In one case, we predict that a stem and loop just upstream of the Shine-Dalgarno sequence sterically hinders the binding of the ribosome to the mRNA.
D001483:Base Sequence; D001665:Binding Sites; D002874:Chromosome Mapping; D004269:DNA, Bacterial / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics*; D007763:Lac Operon; D008074:Lipoproteins / Q000235:genetics; D009154:Mutation; D010957:Plasmids; D014176:Protein Biosynthesis; D012270:Ribosomes
D004269:DNA, Bacterial; D008074:Lipoproteins