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A tRNA aminoacylation system for non-natural amino acids based on a programmable ribozyme

Nature Biotechnology volume 20, pages 723–728 (2002)Cite this article

Abstract

The ability to recognize tRNA identities is essential to the function of the genetic coding system. In translation aminoacyl-tRNA synthetases (ARSs) recognize the identities of tRNAs and charge them with their cognate amino acids. We show that an in vitro–evolved ribozyme can also discriminate between specific tRNAs, and can transfer amino acids to the 3′ ends of cognate tRNAs. The ribozyme interacts with both the CCA-3′ terminus and the anticodon loop of tRNAfMet, and its tRNA specificity is controlled by these interactions. This feature allows us to program the selectivity of the ribozyme toward specific tRNAs, and therefore to tailor effective aminoacyl-transfer catalysts. This method potentially provides a means of generating aminoacyl tRNAs that are charged with non-natural amino acids, which could be incorporated into proteins through cell-free translation.

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Figure 1: Strategy for the isolation of an acyl-transferase ribozyme that recognizes a specific tRNA.
Figure 2: Properties of the BC28 ribozyme.
Figure 3: Specificity of BC28 toward various tRNA substrates.
Figure 4: Programming tRNA specificity into the ribozyme.

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Acknowledgements

We thank all members of the Suga laboratory for helpful discussions, particularly N. Lee for synthesis of substrates. D.R.W.H. is a grateful recipient of a Royal Society–Fulbright Postdoctoral Fellowship (United Kingdom). This work was supported by US National Institutes of Health and National Science Foundation awards to H.S.

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  1. Department of Chemistry, University at Buffalo, State University of New York, Buffalo, 14260-3000, NY

    Yoshitaka Bessho, David R.W. Hodgson & Hiroaki Suga

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  1. Yoshitaka Bessho
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  3. Hiroaki Suga
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Correspondence to Hiroaki Suga.

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Bessho, Y., Hodgson, D. & Suga, H. A tRNA aminoacylation system for non-natural amino acids based on a programmable ribozyme. Nat Biotechnol 20, 723–728 (2002). https://doi.org/10.1038/nbt0702-723

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