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Single nucleotide translation without ribosomes

Abstract

The translation of messenger RNA sequences into polypeptide sequences according to the genetic code is central to life. How this process, which relies on the ribosomal machinery, arose from much simpler precursors is unclear. Here, we demonstrate that single nucleotides charged with an amino acid couple with amino acids linked to the 5′-terminus of an RNA primer in reactions directed by the nucleotides of an RNA template in dilute aqueous solution at 0 °C. When a mixture of U-Val, A-Gly and G-Leu competed for coupling to Gly-RNA, base pairing dictated which dipeptide sequence formed preferentially. The resulting doubly anchored dipeptides can retain their link to the primer for further extension or can be fully released under mild acidic conditions. These results show that a single-nucleotide-based form of translation exists that requires no more than oligoribonucleotides and anchored amino acids.

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Fig. 1: Translation in the cell and single-nucleotide translation presented in this work.
Fig. 2: Template effect in dipeptide formation.
Fig. 3: The RNA template determines which amino acid is coupled to produce a dipeptide.
Fig. 4: Release of a free peptide at the end of translation.
Fig. 5: Extension of dipeptido RNA to tripeptidoyl RNA, as detected by MALDI-TOF mass spectrometry after 45 h at 0 °C.

Data availability

The data generated and analysed during the current project are provided in this paper and its Supplementary Information. Supplementary data, materials and methods are presented in the Supplementary Information. Source data are provided with this paper.

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Acknowledgements

We thank D. Pfeffer for help with high-performance liquid chromatography, H. Griesser for reading and commenting on a draft of the manuscript, E. Kervio and S. Lorenz for discussions, D. Göhringer for technical assistance and C. Guttroff for measuring ESI mass spectra. This work was funded by Deutsche Forschungsgemeinschaft (project-ID 364653263-TRR 235) and the Volkswagen Foundation (grant no. Az 92 768) (to C.R.).

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Contributions

B.J., P.T. and D.J. performed the experiments, C.R. devised and supervised the project. All authors analysed the data. All authors wrote the manuscript and all authors reviewed and commented on the manuscript.

Corresponding author

Correspondence to Clemens Richert.

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The authors declare no competing interests.

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Peer review information Nature Chemistry thanks Thomas Carell and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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Supplementary information

Supplementary Information

Materials and methods, general protocols for synthesis, analytical data, Figs. 1–68 and Tables 1–6.

Source data

Source Data Fig. 2

Source data for Fig. 2c.

Source Data Fig. 3

Source data for Fig. 3b.

Source Data Fig. 5

Source data for Fig. 5.

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Jash, B., Tremmel, P., Jovanovic, D. et al. Single nucleotide translation without ribosomes. Nat. Chem. 13, 751–757 (2021). https://doi.org/10.1038/s41557-021-00749-4

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