ArticleRNA (New York, N.Y.)2010
Missense suppressor mutations in 16S rRNA reveal the importance of helices h8 and h14 in aminoacyl-tRNA selection.
Article in RNA (New York, N.Y.), 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 44 papers.
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Who cites it
44 citing papers in PubMed, 60 citations in OpenAlex.
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- Multiscale Modeling of the Bacterial Ribosome to Identify Potential Peptide Modulators and Their Allosteric Effects.Biochemistry · 2026Article
- Contribution of an alternative 16S rRNA helix to biogenesis of the 30S subunit of the ribosome.RNA (New York, N.Y.) · 2024Article
- Engineered mRNA-ribosome fusions for facile biosynthesis of selenoproteins.Proceedings of the National Academy of Sciences of the United States of America · 2024Article
- Genome-wide screening reveals metabolic regulation of stop-codon readthrough by cyclic AMP.Nucleic acids research · 2023Article
- Roles of the leader-trailer helix and antitermination complex in biogenesis of the 30S ribosomal subunit.Nucleic acids research · 2023Article
- Cryo-EM reconstruction of the human 40S ribosomal subunit at 2.15 Å resolution.Nucleic acids research · 2023Article
- Bridging ribosomal synthesis to cell growth through the lens of kinetics.Biophysical journal · 2023Article
- Our current understanding of the toxicity of altered mito-ribosomal fidelity during mitochondrial protein synthesis: What can it tell us about human disease?Frontiers in physiology · 2023Review
- Structural analysis of mitochondrial rRNA gene variants identified in patients with deafness.Frontiers in physiology · 2023Article
- The universally conserved nucleotides of the small subunit ribosomal RNAs.RNA (New York, N.Y.) · 2022Article
- Regulation of translation by site-specific ribosomal RNA methylation.Nature structural & molecular biology · 2021Article
- Article
- Mutational characterization and mapping of the 70S ribosome active site.Nucleic acids research · 2020Article
- Fail-safe genetic codes designed to intrinsically contain engineered organisms.Nucleic acids research · 2019Article
- Optimal translational fidelity is critical for Salmonella virulence and host interactions.Nucleic acids research · 2019Article
- Ribosomal ambiguity (ram) mutations promote the open (off) to closed (on) transition and thereby increase miscoding.Nucleic acids research · 2019Article
- Decoding on the ribosome depends on the structure of the mRNA phosphodiester backbone.Proceedings of the National Academy of Sciences of the United States of America · 2018Article
- Translational fidelity and mistranslation in the cellular response to stress.Nature microbiology · 2017Review
- Modular Organization of Residue-Level Contacts Shapes the Selection Pressure on Individual Amino Acid Sites of Ribosomal Proteins.Genome biology and evolution · 2017Article
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
Abstract
The molecular basis of the induced-fit mechanism that determines the fidelity of protein synthesis remains unclear. Here, we isolated mutations in 16S rRNA that increase the rate of miscoding and stop codon read-through. Many of the mutations clustered along interfaces between the 30S shoulder domain and other parts of the ribosome, strongly implicating shoulder movement in the induced-fit mechanism of decoding. The largest subset of mutations mapped to helices h8 and h14. These helices interact with each other and with the 50S subunit to form bridge B8. Previous cryo-EM studies revealed a contact between h14 and the switch 1 motif of EF-Tu, raising the possibility that h14 plays a direct role in GTPase activation. To investigate this possibility, we constructed both deletions and insertions in h14. While ribosomes harboring a 2-base-pair (bp) insertion in h14 were completely inactive in vivo, those containing a 2-bp deletion retained activity but were error prone. In vitro, the truncation of h14 accelerated GTP hydrolysis for EF-Tu bearing near-cognate aminoacyl-tRNA, an effect that can largely account for the observed miscoding in vivo. These data show that h14 does not help activate EF-Tu but instead negatively controls GTP hydrolysis by the factor. We propose that bridge B8 normally acts to counter inward rotation of the shoulder domain; hence, mutations in h8 and h14 that compromise this bridge decrease the stringency of aminoacyl-tRNA selection.
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