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Bases in the anticodon loop of tRNAAlaGGC prevent misreading

Nature Structural & Molecular Biology volume 16, pages 353–358 (2009)Cite this article

Abstract

The bases at positions 32 and 38 in the tRNA anticodon loop are known to have a specific conservation depending upon the anticodon triplets. Here we report that evolutionarily conserved pairs of bases at positions 32 and 38 in tRNAAlaGGC prevent misreading of a near-cognate valine codon, GUC. The tRNAAlaGGC molecules with the conserved A32-U38 and C32-G38 pairs do not read GUC, whereas those with three representative nonconserved pairs, U32-U38, U32-A38 and C32-A38, direct the misincorporation of alanine at this valine codon into the peptide chain. Overexpression of the nonconserved tRNAAlaGGC in Escherichia coli is toxic and prevents cell growth. These results suggested that the bases at positions 32 and 38 in tRNAAlaGGC evolved to preserve the fidelity of the cognate codon reading.

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Figure 1: Structure of wild-type tRNAAlaGGC and its variants
Figure 2: Decoding efficiency of the GCC codon by tRNAAlaGGC with the conserved or nonconserved 32-38 pair.
Figure 3: Influence of the sequence variation of the 32-38 pair in tRNAAlaGGC on misreading of GUC codon.
Figure 4: Overexpression of the conserved or nonconserved tRNAAlaGGC in E. coli (BL21).

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Acknowledgements

We thank O.C. Uhlenbeck and S. Ledoux for their invaluable discussion. This work was supported by grants from the Japan Society for the Promotion of Science (JSPS) Grants-in-Aid for Scientific Research (S) (16101007) to H.S., a Young Scientists (A) (20681022) to H.M., a JSPS Fellowship (19-1722) to A.O., a research and development project of the Industrial Science and Technology Program in the New Energy and Industrial Technology Development Organization (NEDO) to H.S., the Industrial Technology Research Grant Program in NEDO (05A02513a) to H.M., and the Takeda Science Foundation.

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Authors and Affiliations

  1. Research Center for Advanced Science and Technology, University of Tokyo, Meguro-ku, Tokyo, Japan

    Hiroshi Murakami & Hiroaki Suga

  2. Department of Chemistry and Biotechnology, Graduate School of Engineering, University of Tokyo, Bunkyo-ku, Tokyo, Japan

    Atsushi Ohta & Hiroaki Suga

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  1. Hiroshi Murakami
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  2. Atsushi Ohta
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This study was designed by H.M., A.O. and H.S.; all of the experiments were performed by H.M.; the paper was written by H.M. and H.S.

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Correspondence to Hiroshi Murakami or Hiroaki Suga.

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Murakami, H., Ohta, A. & Suga, H. Bases in the anticodon loop of tRNAAlaGGC prevent misreading. Nat Struct Mol Biol 16, 353–358 (2009). https://doi.org/10.1038/nsmb.1580

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