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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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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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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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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DOI: https://doi.org/10.1038/nsmb.1580
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