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Quality control by the ribosome following peptide bond formation

Nature volume 457, pages 161–166 (2009)Cite this article

Abstract

The overall fidelity of protein synthesis has been thought to rely on the combined accuracy of two basic processes: the aminoacylation of transfer RNAs with their cognate amino acid by the aminoacyl-tRNA synthetases, and the selection of cognate aminoacyl-tRNAs by the ribosome in cooperation with the GTPase elongation factor EF-Tu. These two processes, which together ensure the specific acceptance of a correctly charged cognate tRNA into the aminoacyl (A) site, operate before peptide bond formation. Here we report the identification of an additional mechanism that contributes to high fidelity protein synthesis after peptidyl transfer, using a well-defined in vitro bacterial translation system. In this retrospective quality control step, the incorporation of an amino acid from a non-cognate tRNA into the growing polypeptide chain leads to a general loss of specificity in the A site of the ribosome, and thus to a propagation of errors that results in abortive termination of protein synthesis.

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Figure 1: Unusual behaviour by RF2 after a miscoding event.
Figure 2: Abortive termination reaction is stimulated by the class II RF3 and is general for all P-site mismatches.
Figure 3: A single miscoding event promotes iterated errors in tRNA selection.
Figure 4: Iterated miscoding results in doubly mismatched complexes, where release catalysis is dramatically promoted.
Figure 5: An initial miscoding event results in an overall drop in yield of full-length peptides.

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Acknowledgements

We thank B. Cormack, A. Nahvi, A. Buskirk and R. Reed for helpful comments on the manuscript and members of the laboratory for useful discussions. This work was supported by the National Institutes of Health with salary support from Howard Hughes Medical Institute.

Author Contributions H.S.Z. and R.G. designed the experiments and wrote the manuscript. H.S.Z. performed the experiments.

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  1. Department of Molecular Biology and Genetics, Howard Hughes Medical Institute, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA,

    Hani S. Zaher & Rachel Green

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  1. Hani S. Zaher
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Correspondence to Rachel Green.

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Zaher, H., Green, R. Quality control by the ribosome following peptide bond formation. Nature 457, 161–166 (2009). https://doi.org/10.1038/nature07582

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