Abstract
The Watson-Crick base pairs between the 3′-terminal end of tRNAs and ribosomal RNA in the peptidyl transferase center are universally conserved. Here, we report that the introduction of compensatory mutations to Escherichia coli RNAs in this site leads to an orthogonal system independent of the wild-type counterpart, as demonstrated via the production of two peptide sequences from a single mRNA. This work thus identifies a new way to reprogram the genetic code.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Bashan, A. et al. Mol. Cell 11, 91–102 (2003).
Hansen, J.L., Schmeing, T.M., Moore, P.B. & Steitz, T.A. Proc. Natl. Acad. Sci. USA 99, 11670–11675 (2002).
Nissen, P., Hansen, J., Ban, N., Moore, P.B. & Steitz, T.A. Science 289, 920–930 (2000).
Steitz, T.A. Nat. Rev. Mol. Cell Biol. 9, 242–253 (2008).
Moazed, D. & Noller, H.F. Proc. Natl. Acad. Sci. USA 88, 3725–3728 (1991).
Samaha, R.R., Green, R. & Noller, H.F. Nature 377, 309–314 (1995).
Lescoute, A. & Westhof, E. Nucleic Acids Res. 34, 6587–6604 (2006).
Green, R., Switzer, C. & Noller, H.F. Science 280, 286–289 (1998).
Kim, D.F. & Green, R. Mol. Cell 4, 859–864 (1999).
Cavarelli, J. & Moras, D. FASEB J. 7, 79–86 (1993).
Ruff, M. et al. Science 252, 1682–1689 (1991).
Schulman, L.H. & Pelka, H. Biochemistry 16, 4256–4265 (1977).
Liu, M. & Horowitz, J. Proc. Natl. Acad. Sci. USA 91, 10389–10393 (1994).
Zhou, X.L. et al. Nucleic Acids Res. 39, 8857–8868 (2011).
Murakami, H., Ohta, A., Ashigai, H. & Suga, H. Nat. Methods 3, 357–359 (2006).
Ohuchi, M., Murakami, H. & Suga, H. Curr. Opin. Chem. Biol. 11, 537–542 (2007).
Xiao, H., Murakami, H., Suga, H. & Ferré-D'Amaré, A.R. Nature 454, 358–361 (2008).
Saito, H., Kourouklis, D. & Suga, H. EMBO J. 20, 1797–1806 (2001).
Ohta, A., Murakami, H., Higashimura, E. & Suga, H. Chem. Biol. 14, 1315–1322 (2007).
Kawakami, T. et al. Nat. Chem. Biol. 5, 888–890 (2009).
Goto, Y., Katoh, T. & Suga, H. Nat. Protoc. 6, 779–790 (2011).
Youngman, E.M. & Green, R. Methods 36, 305–312 (2005).
Sako, Y., Morimoto, J., Murakami, H. & Suga, H. J. Am. Chem. Soc. 130, 7232–7234 (2008).
Kao, C., Zheng, M. & Rüdisser, S. RNA 5, 1268–1272 (1999).
Doi, Y., Ohtsuki, T., Shimizu, Y., Ueda, T. & Sisido, M. J. Am. Chem. Soc. 129, 14458–14462 (2007).
Acknowledgements
We thank R. Green for the gift of a ribosome construct linked to the MS2 tag. This work was supported by Japan Society for the Promotion of Science (JSPS) Grants-in-Aid for Specially Promoted Research (21000005) and the Japan Science and Technology Agency (JST) Core Research for Evolutional Science and Technology (CREST) of Molecular Technology to H.S. N.T. (24-8188) and G.H. (21-9361) are supported by Grants-in-Aid for JSPS Fellows. T.K. is supported by a JSPS Grant-in-Aid for Young Scientists (A) (24681047) and a JST Rising Star Award of Molecular Technology.
Author information
Authors and Affiliations
Contributions
N.T. and G.H. conducted biochemical and chemical studies, and T.K. and H.S. supervised the research. All of the authors contributed to writing the manuscript.
Corresponding author
Ethics declarations
Competing interests
The authors declare no competing financial interests.
Supplementary information
Supplementary Text and Figures
Supplementary Results. (PDF 6948 kb)
Rights and permissions
About this article
Cite this article
Terasaka, N., Hayashi, G., Katoh, T. et al. An orthogonal ribosome-tRNA pair via engineering of the peptidyl transferase center. Nat Chem Biol 10, 555–557 (2014). https://doi.org/10.1038/nchembio.1549
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/nchembio.1549
This article is cited by
-
Escherichia coli methionine-tRNAi/methionyl tRNA synthetase pairs induced protein initiation of interest (PII) expression
Applied Biological Chemistry (2022)
-
Controlling orthogonal ribosome subunit interactions enables evolution of new function
Nature (2018)
-
Ribosomal synthesis and folding of peptide-helical aromatic foldamer hybrids
Nature Chemistry (2018)
-
Next-generation biocontainment systems for engineered organisms
Nature Chemical Biology (2018)
-
Expanding and reprogramming the genetic code
Nature (2017)