title: Mutational robustness of 16S ribosomal RNA, shown by experimental horizontal gene transfer in Escherichia coli.
abstract: The bacterial ribosome consists of three rRNA molecules and 57 proteins and plays a crucial role in translating mRNA-encoded information into proteins. Because of the ribosome's structural and mechanistic complexity, it is believed that each ribosomal component coevolves to maintain its function. Unlike 5S rRNA, 16S and 23S rRNAs appear to lack mutational robustness, because they form the structural core of the ribosome. However, using Escherichia coli Δ7 (null mutant of operons) as a host, we have recently shown that an active hybrid ribosome whose 16S rRNA has been specifically substituted with that from non-E. coli bacteria can be reconstituted in vivo. To investigate the mutational robustness of 16S rRNA and the structural basis for its functionality, we used a metagenomic approach to screen for 16S rRNA genes that complement the growth of E. coli Δ7. Various functional genes were obtained from the Gammaproteobacteria and Betaproteobacteria lineages. Despite the large sequence diversity (80.9-99.0% identity with E. coli 16S rRNA) of the functional 16S rRNA molecules, the doubling times (DTs) of each mutant increased only modestly with decreasing sequence identity (average increase in DT, 4.6 s per mutation). The three-dimensional structure of the 30S ribosome showed that at least 40.7% (628/1,542) of the nucleotides were variable, even at ribosomal protein-binding sites, provided that the secondary structures were properly conserved. Our results clearly demonstrate that 16S rRNA functionality largely depends on the secondary structure but not on the sequence itself.
authors.affiliation: Bioproduction Research Institute,  National Institute of Advanced Industrial Science and Technology, Sapporo 062-8517,  Japan.
authors.forename: Kei
authors.initials: K
authors.lastname: Kitahara
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Yasutake
authors.forename: Kentaro
authors.initials: K
authors.lastname: Miyazaki
mesh_terms: D001483:Base Sequence; D004926:Escherichia coli / Q000235:genetics*; D018014:Gene Transfer Techniques; D022761:Gene Transfer, Horizontal / Q000235:genetics; D005798:Genes, Bacterial / Q000235:genetics; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics*; D009690:Nucleic Acid Conformation; D009711:Nucleotides / Q000235:genetics; D010802:Phylogeny; D011485:Protein Binding; D012329:RNA, Bacterial / Q000737:chemistry / Q000235:genetics*; D012336:RNA, Ribosomal, 16S / Q000737:chemistry / Q000235:genetics*; D012269:Ribosomal Proteins / Q000378:metabolism
chemical_list: D009711:Nucleotides; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S; D012269:Ribosomal Proteins