title: Coevolution of RNA helix stability and Shine-Dalgarno complementarity in a translational start region.
abstract: The initiation region of the coat-protein gene of RNA bacteriophage MS2 adopts a well-defined hairpin structure with the start codon occupying the loop position, while the Shine-Dalgarno (SD) sequence is part of the stem. In a previous study, we introduced mutations in this hairpin that changed its thermodynamic stability. The resulting phages evolved to regain the wild-type stability by second-site compensatory substitutions. Neither the original nor the suppressor mutations were in the SD region. In the present analysis, we have made changes in the SD region that shorten or extend its complementarity to the 3' end of 16S rRNA and monitored their evolution to a stable pseudorevertant species. Phages in which the SD complementarity was decreased evolved an initiator hairpin of lower stability than wild type while those in which the complementarity was extended evolved a hairpin with an increased stability. We conclude that weaker SD sequences still allow maximal translation if the secondary structure of the ribosome-landing site is destabilized accordingly. Alternatively, translation-initiation regions with a stronger secondary structure still allow maximal expression, if the SD complementarity is extended. These findings support a previously published model in which the SD interaction helps the ribosome to melt the structure in a translation-initiation region.
authors.affiliation: Department of Biochemistry, Leiden Institute of Chemistry, Gorlaeus Laboratories, University of Leiden, The Netherlands.
authors.forename: R C
authors.initials: RC
authors.lastname: Olsthoorn
authors.forename: S
authors.initials: S
authors.lastname: Zoog
authors.forename: J
authors.initials: J
authors.lastname: van Duin
mesh_terms: D001483:Base Sequence; D001665:Binding Sites; D005075:Biological Evolution; D002213:Capsid / Q000235:genetics*; D036022:Capsid Proteins; D003062:Codon; D017909:Levivirus / Q000235:genetics*; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009154:Mutation; D009690:Nucleic Acid Conformation; D017354:Point Mutation; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000737:chemistry* / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000737:chemistry; D012367:RNA, Viral / Q000737:chemistry* / Q000235:genetics; D016601:RNA-Binding Proteins; D011994:Recombinant Proteins / Q000737:chemistry; D012045:Regulatory Sequences, Nucleic Acid; D012270:Ribosomes / Q000378:metabolism
chemical_list: D036022:Capsid Proteins; D003062:Codon; D012333:RNA, Messenger; D012336:RNA, Ribosomal, 16S; D012367:RNA, Viral; D016601:RNA-Binding Proteins; D011994:Recombinant Proteins