ArticleProceedings of the National Academy of Sciences of the United States of America2007
A natural genetic code expansion cassette enables transmissible biosynthesis and genetic encoding of pyrrolysine.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2007. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 39 papers.
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Who cites it
39 citing papers in PubMed, 83 citations in OpenAlex.
- Pyrrolysine supply and demand shapes the translational landscape inProceedings of the National Academy of Sciences of the United States of America · 2026Article
- Methanogenic archaea encoding Pyrrolysine maintain ambiguous amber codon usage.Proceedings of the National Academy of Sciences of the United States of America · 2025Article
- Genetic Code Expansion: Recent Developments and Emerging Applications.Chemical reviews · 2025Review
- Engineering Pyrrolysine Systems for Genetic Code Expansion and Reprogramming.Chemical reviews · 2024Review
- Cellular Site-Specific Incorporation of Noncanonical Amino Acids in Synthetic Biology.Chemical reviews · 2024Review
- Advances in Biosynthesis of Non-Canonical Amino Acids (ncAAs) and the Methods of ncAAs Incorporation into Proteins.Molecules (Basel, Switzerland) · 2023Review
- Update of the Pyrrolysyl-tRNA Synthetase/tRNAJournal of bacteriology · 2023Review
- Recoding of stop codons expands the metabolic potential of two novel Asgardarchaeota lineages.ISME communications · 2021Article
- Improved pyrrolysine biosynthesis through phage assisted non-continuous directed evolution of the complete pathway.Nature communications · 2021Article
- Programmed Deviations of Ribosomes From Standard Decoding in Archaea.Frontiers in microbiology · 2021Review
- Plasticity and Constraints of tRNA Aminoacylation Define Directed Evolution of Aminoacyl-tRNA Synthetases.International journal of molecular sciences · 2019Review
- Facile Method of Protein PEGylation by a Mono-Ion Complex.ACS omega · 2017Article
- Article
- Engineering microbial hosts for production of bacterial natural products.Natural product reports · 2016Review
- Pyrrolysyl-tRNA synthetase, an aminoacyl-tRNA synthetase for genetic code expansion.Croatica chemica acta. Arhiv za kemiju · 2016Article
- Reducing the genetic code induces massive rearrangement of the proteome.Proceedings of the National Academy of Sciences of the United States of America · 2014Article
- Pyrrolysyl-tRNA synthetase: an ordinary enzyme but an outstanding genetic code expansion tool.Biochimica et biophysica acta · 2014Review
- Unique characteristics of the pyrrolysine system in the 7th order of methanogens: implications for the evolution of a genetic code expansion cassette.Archaea (Vancouver, B.C.) · 2014Article
- Multiple site-selective insertions of noncanonical amino acids into sequence-repetitive polypeptides.Chembiochem : a European journal of chemical biology · 2013Article
- Carbon source-dependent expansion of the genetic code in bacteria.Proceedings of the National Academy of Sciences of the United States of America · 2012Article
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Authors and funding
7 authors at 2 institutions in 2 countries.
Funding
Abstract
Pyrrolysine has entered natural genetic codes by the translation of UAG, a canonical stop codon. UAG translation as pyrrolysine requires the pylT gene product, an amber-decoding tRNA(Pyl) that is aminoacylated with pyrrolysine by the pyrrolysyl-tRNA synthetase produced from the pylS gene. The pylTS genes form a gene cluster with pylBCD, whose functions have not been investigated. The pylTSBCD gene order is maintained not only in methanogenic Archaea but also in a distantly related Gram-positive Bacterium, indicating past horizontal gene transfer of all five genes. Here we show that lateral transfer of pylTSBCD introduces biosynthesis and genetic encoding of pyrrolysine into a naïve organism. PylS-based assays demonstrated that pyrrolysine was biosynthesized in Escherichia coli expressing pylBCD from Methanosarcina acetivorans. Production of pyrrolysine did not require tRNA(Pyl) or PylS. However, when pylTSBCD were coexpressed with mtmB1, encoding the methanogen monomethylamine methyltransferase, UAG was translated as pyrrolysine to produce recombinant monomethylamine methyltransferase. Expression of pylTSBCD also suppressed an amber codon introduced into the E. coli uidA gene. Strains lacking one of the pylBCD genes did not produce pyrrolysine or translate UAG as pyrrolysine. These results indicated that pylBCD gene products biosynthesize pyrrolysine using metabolites common to Bacteria and Archaea and, furthermore, that the pyl gene cluster represents a "genetic code expansion cassette," previously unprecedented in natural organisms, whose transfer allows an existing codon to be translated as a novel endogenously synthesized free amino acid. Analogous cassettes may have served similar functions for other amino acids during the evolutionary expansion of the canonical genetic code.
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