ArticleProceedings of the National Academy of Sciences of the United States of America2004
Isolation of antibiotic resistance mutations in the rRNA by using an in vitro selection system.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2004. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 21 papers.
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21 citing papers in PubMed, 46 citations in OpenAlex.
- Autonomous ribosome biogenesis in vitro.Nature communications · 2025Article
- Long-term exposure to food-grade disinfectants causes cross-resistance to antibiotics in Salmonella enterica serovar Typhimurium strains with different antibiograms and sequence types.Antimicrobial resistance and infection control · 2023Article
- Mapping the in vivo fitness landscape of a tethered ribosome.Science advances · 2023Article
- Community science designed ribosomes with beneficial phenotypes.Nature communications · 2023Article
- Computationally-guided design and selection of high performing ribosomal active site mutants.Nucleic acids research · 2022Article
- Targeted editing and evolution of engineered ribosomes in vivo by filtered editing.Nature communications · 2022Article
- Orthogonal translation enables heterologous ribosome engineering in E. coli.Nature communications · 2021Article
- Ribosome-mediated polymerization of long chain carbon and cyclic amino acids into peptides in vitro.Nature communications · 2020Article
- A fully orthogonal system for protein synthesis in bacterial cells.Nature communications · 2020Article
- In vitro ribosome synthesis and evolution through ribosome display.Nature communications · 2020Article
- Strategies for in vitro engineering of the translation machinery.Nucleic acids research · 2020Review
- Single enzyme RT-PCR of full-length ribosomal RNA.Synthetic biology (Oxford, England) · 2020Article
- Expanding the Scope of Protein Synthesis Using Modified Ribosomes.Journal of the American Chemical Society · 2019Article
- Repurposing ribosomes for synthetic biology.Current opinion in chemical biology · 2017Review
- Resistance mutations generate divergent antibiotic susceptibility profiles against translation inhibitors.Proceedings of the National Academy of Sciences of the United States of America · 2016Article
- Expedited quantification of mutant ribosomal RNA by binary deoxyribozyme (BiDz) sensors.RNA (New York, N.Y.) · 2015Article
- The impact of transcriptional tuning on in vitro integrated rRNA transcription and ribosome construction.Nucleic acids research · 2014Article
- Tools for characterizing bacterial protein synthesis inhibitors.Antimicrobial agents and chemotherapy · 2013Article
- In vitro integration of ribosomal RNA synthesis, ribosome assembly, and translation.Molecular systems biology · 2013Article
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2 authors at 1 institution in 1 country.
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No grant is acknowledged in the PubMed record.
Abstract
Genetic, biochemical, and structural data support an essential role for the ribosomal RNA in all steps of the translation process. Although in vivo genetic selection techniques have been used to identify mutations in the rRNAs that result in various miscoding phenotypes and resistance to known ribosome-targeted antibiotics, these are limited because the resulting mutant ribosomes must be only marginally disabled if they are able to support growth of the cell. Furthermore, in vivo, it is not possible to control the environment in precise ways that might allow for the isolation of certain types of rRNA variants. To overcome these limitations, we have developed an in vitro selection system for the isolation of functionally competent ribosomal particles from populations containing variant rRNAs. Here, we describe this system and present an example of its application to the selection of antibiotic resistance mutations. From a pool of 4,096 23S rRNA variants, a double mutant (A2058U/A2062G) was isolated after iteration of the selection process. This mutant was highly resistant to clindamycin in in vitro translation reactions and yet was not viable in Escherichia coli. These data establish that this system has the potential to identify mutations in the rRNA not readily accessed by comparable in vivo systems, thus allowing for more exhaustive ribosomal genetic screens.
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