Innate immune and chemically triggered oxidative stress modifies translational fidelity (original) (raw)

Nature volume 462, pages 522–526 (2009)Cite this article

Abstract

Translational fidelity, essential for protein and cell function, requires accurate transfer RNA (tRNA) aminoacylation. Purified aminoacyl-tRNA synthetases exhibit a fidelity of one error per 10,000 to 100,000 couplings1,2. The accuracy of tRNA aminoacylation in vivo is uncertain, however, and might be considerably lower3,4,5,6. Here we show that in mammalian cells, approximately 1% of methionine (Met) residues used in protein synthesis are aminoacylated to non-methionyl-tRNAs. Remarkably, Met-misacylation increases up to tenfold upon exposing cells to live or non-infectious viruses, toll-like receptor ligands or chemically induced oxidative stress. Met is misacylated to specific non-methionyl-tRNA families, and these Met-misacylated tRNAs are used in translation. Met-misacylation is blocked by an inhibitor of cellular oxidases, implicating reactive oxygen species (ROS) as the misacylation trigger. Among six amino acids tested, tRNA misacylation occurs exclusively with Met. As Met residues are known to protect proteins against ROS-mediated damage7, we propose that Met-misacylation functions adaptively to increase Met incorporation into proteins to protect cells against oxidative stress. In demonstrating an unexpected conditional aspect of decoding mRNA, our findings illustrate the importance of considering alternative iterations of the genetic code.

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Gene Expression Omnibus

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The microarray platforms are deposited in the National Center for Biotechnology Information (NCBI) Gene Expression Omnibus (GEO) database under accession numbers GPL9427 and GPL9428.

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Acknowledgements

The authors are grateful to D. Klinman for his gift of CpG oligonucleotides and advice, A. Schilling for supervision and advice on mass spectrometry experiments, and C. Nicchitta, T. Pierson, P. Cluzel, R. Levine, A. Iwasaki and S. Amigorena for insight and advice. This work was supported by the Division of Intramural Research, the National Institute of Allergy and Infectious Diseases, and by National Institutes of Health extramural pilot projects.

Author Contributions N.N., J.M.G., A.D., K.A.D., R.B.J., J.R.S., D.B., E.M.E, M.R.R., J.S.B., A.E., B.D., S.D., H.D.H., P.B. and T.P. designed and performed experiments, and analysed data. J.S.G. generated genetic constructs. J.R.B. and J.W.Y designed experiments and analysed data. J.W.Y. and T.P. conceived the project and wrote the paper. J.W.Y. and T.P. contributed equally to this paper.

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Author notes

  1. Nir Netzer, Jeffrey M. Goodenbour, Jonathan W. Yewdell and Tao Pan: These authors contributed equally to this work.

Authors and Affiliations

  1. Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland 20892, USA ,
    Nir Netzer, Alexandre David, James S. Gibbs, Alan Embry, Brian Dolan, Suman Das, Heather D. Hickman, Peter Berglund, Jack R. Bennink & Jonathan W. Yewdell
  2. Department of Human Genetics,,
    Jeffrey M. Goodenbour
  3. Department of Biochemistry and Molecular Biology,,
    Kimberly A. Dittmar & Tao Pan
  4. Ben May Institute,,
    Richard B. Jones, Eva M. Eves & Marsha R. Rosner
  5. Department of Medicine, University of Chicago, Chicago, Illinois 60637, USA,
    Jeffrey R. Schneider & David Boone

Authors

  1. Nir Netzer
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  2. Jeffrey M. Goodenbour
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  3. Alexandre David
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  4. Kimberly A. Dittmar
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  5. Richard B. Jones
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  6. Jeffrey R. Schneider
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  7. David Boone
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  8. Eva M. Eves
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  9. Marsha R. Rosner
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  10. James S. Gibbs
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  11. Alan Embry
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  12. Brian Dolan
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  13. Suman Das
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  14. Heather D. Hickman
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  15. Peter Berglund
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  16. Jack R. Bennink
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  17. Jonathan W. Yewdell
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  18. Tao Pan
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Corresponding authors

Correspondence toJonathan W. Yewdell or Tao Pan.

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Netzer, N., Goodenbour, J., David, A. et al. Innate immune and chemically triggered oxidative stress modifies translational fidelity.Nature 462, 522–526 (2009). https://doi.org/10.1038/nature08576

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Editorial Summary

Mutation by stealth

For cells to function properly the process of translating RNA messengers into proteins needs to be accurate, on the whole. Yet work in HeLa cells now shows that about 1% of methionine residues used in protein synthesis are aminoacylated to 'textbook-incorrect' tRNAs. Surprisingly, the proportion of Met-misacylated tRNAs increases significantly when cells are under stress through viral infection or treatment with viral or bacterial Toll-like receptor ligands. Tests with other amino acids indicate that the phenomenon is limited to Met, and as Met residues are known to protect proteins against damage from reactive oxygen species, one possibility is that Met-misacylation is a natural protective response to cellular stress.