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Ribozyme-catalyzed tRNA aminoacylation

Nature Structural Biology volume 7, pages 28–33 (2000)Cite this article

Abstract

The RNA world hypothesis implies that coded protein synthesis evolved from a set of ribozyme catalyzed acyl-transfer reactions, including those of aminoacyl-tRNA synthetase ribozymes. We report here that a bifunctional ribozyme generated by directed in vitro evolution can specifically recognize an activated glutaminyl ester and aminoacylate a targeted tRNA, via a covalent aminoacyl-ribozyme intermediate. The ribozyme consists of two distinct catalytic domains; one domain recognizes the glutamine substrate and self-aminoacylates its own 5'-hydroxyl group, and the other recognizes the tRNA and transfers the aminoacyl group to the 3'-end. The interaction of these domains results in a unique pseudoknotted structure, and the ribozyme requires a change in conformation to perform the sequential aminoacylation reactions. Our result supports the idea that aminoacyl-tRNA synthetase ribozymes could have played a key role in the evolution of the genetic code and RNA-directed translation.

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Figure 1: Secondary structure of acyl-transferase ribozyme (ATRib) and schematic representation of a ATRib-catalyzed aminoacylation of acceptor RNA.
Figure 2: Aminoacylation of tRNA catalyzed by ATRib.
Figure 3: In vitro evolution of aaRS-like ribozymes.
Figure 4: Sequence alignment of active clones in pool 12 DNA and amino acid specificity of clone AD02.
Figure 5: Structure and function of the ribozyme.

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Acknowledgements

We thank the NMR facility in the Department of Chemistry, the CAMBI nucleic acid facility, and the Phosphorimager facility in the Department of Biological Sciences. We thank M. Hollingsworth and P. Gollnick for critical reading the manuscript. Y.B. is supported by a Research Scientist Abroad Fellowship sponsored by the Japanese Ministry of Education. This work was supported by the State University of New York at Buffalo Start-up Fund and a PRF-ACS grant (H.S.) and partly by an NIH grant (J.W.S.).

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

    Nick Lee, Yoshitaka Bessho, Kenneth Wei & Hiroaki Suga

  2. Department of Molecular Biology, Massachusetts General Hospital, Boston, 02114, Massachusetts, USA

    Jack W. Szostak

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Correspondence to Hiroaki Suga.

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Lee, N., Bessho, Y., Wei, K. et al. Ribozyme-catalyzed tRNA aminoacylation. Nat Struct Mol Biol 7, 28–33 (2000). https://doi.org/10.1038/71225

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