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An orthogonal ribosome-tRNA pair via engineering of the peptidyl transferase center

Nature Chemical Biology volume 10, pages 555–557 (2014)Cite this article

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Abstract

The Watson-Crick base pairs between the 3′-terminal end of tRNAs and ribosomal RNA in the peptidyl transferase center are universally conserved. Here, we report that the introduction of compensatory mutations to Escherichia coli RNAs in this site leads to an orthogonal system independent of the wild-type counterpart, as demonstrated via the production of two peptide sequences from a single mRNA. This work thus identifies a new way to reprogram the genetic code.

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Figure 1: Base pair interaction of the 3′ end of tRNAs with 23S rRNA.
Figure 2: Translational activity of pairs of ribosome and tRNA mutants.
Figure 3: Simultaneous expression of two distinct peptides from a single mRNA sequence under two artificially programmed genetic codes.

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Acknowledgements

We thank R. Green for the gift of a ribosome construct linked to the MS2 tag. This work was supported by Japan Society for the Promotion of Science (JSPS) Grants-in-Aid for Specially Promoted Research (21000005) and the Japan Science and Technology Agency (JST) Core Research for Evolutional Science and Technology (CREST) of Molecular Technology to H.S. N.T. (24-8188) and G.H. (21-9361) are supported by Grants-in-Aid for JSPS Fellows. T.K. is supported by a JSPS Grant-in-Aid for Young Scientists (A) (24681047) and a JST Rising Star Award of Molecular Technology.

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Author notes

  1. Gosuke Hayashi

    Present address: Present address: Research Center for Advanced Science and Technology, University of Tokyo, Tokyo, Japan.,

  2. Naohiro Terasaka and Gosuke Hayashi: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Chemistry, Graduate School of Science, University of Tokyo, Tokyo, Japan

    Naohiro Terasaka, Gosuke Hayashi, Takayuki Katoh & Hiroaki Suga

  2. Japan Science and Technology Agency, Core Research for Evolutional Science and Technology, University of Tokyo, Tokyo, Japan

    Hiroaki Suga

Authors
  1. Naohiro Terasaka
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  2. Gosuke Hayashi
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  3. Takayuki Katoh
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  4. Hiroaki Suga
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Contributions

N.T. and G.H. conducted biochemical and chemical studies, and T.K. and H.S. supervised the research. All of the authors contributed to writing the manuscript.

Corresponding author

Correspondence to Hiroaki Suga.

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The authors declare no competing financial interests.

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Terasaka, N., Hayashi, G., Katoh, T. et al. An orthogonal ribosome-tRNA pair via engineering of the peptidyl transferase center. Nat Chem Biol 10, 555–557 (2014). https://doi.org/10.1038/nchembio.1549

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