Abstract
Genetic code expansion for unnatural amino acid mutagenesis has, until recently, been limited to cell culture. We demonstrate the site-specific incorporation of unnatural amino acids into proteins in Drosophila melanogaster at different developmental stages, in specific tissues and in a subset of cells within a tissue. This approach provides a foundation for probing and controlling processes in this established metazoan model organism with a new level of molecular precision.
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References
Davis, L. & Chin, J.W. Nat. Rev. Mol. Cell Biol. 13, 168–182 (2012).
Liu, C.C. & Schultz, P.G. Annu. Rev. Biochem. 79, 413–444 (2010).
Gautier, A., Deiters, A. & Chin, J.W. J. Am. Chem. Soc. 133, 2124–2127 (2011).
Gautier, A. et al. J. Am. Chem. Soc. 132, 4086–4088 (2010).
Lemke, E.A., Summerer, D., Geierstanger, B.H., Brittain, S.M. & Schultz, P.G. Nat. Chem. Biol. 3, 769–772 (2007).
Rubin, G.M. & Lewis, E.B. Science 287, 2216–2218 (2000).
Adams, M.D. et al. Science 287, 2185–2195 (2000).
Brand, A.H. & Perrimon, N. Development 118, 401–415 (1993).
Rubin, G.M. & Spradling, A.C. Science 218, 348–353 (1982).
Laski, F.A., Ganguly, S. & Sharp, P.A. Proc. Natl. Acad. Sci. USA 86, 6696–6698 (1989).
Garza, D., Medhora, M.M. & Hartl, D.L. Genetics 126, 625–637 (1990).
Washburn, T. & O'Tousa, J.E. Genetics 130, 585–595 (1992).
Lang, K. et al. Nat. Chem. 4, 298–304 (2012).
Nguyen, D.P. et al. J. Am. Chem. Soc. 131, 8720–8721 (2009).
Nguyen, D.P., Garcia Alai, M.M., Kapadnis, P.B., Neumann, H. & Chin, J.W. J. Am. Chem. Soc. 131, 14194–14195 (2009).
Neumann, H., Peak-Chew, S.Y. & Chin, J.W. Nat. Chem. Biol. 4, 232–234 (2008).
Nguyen, D.P. Unnatural Amino Acid Incorporation via the Orthogonal Pyrrolysyl-tRNA Synthetase–tRNACUA Pair. PhD thesis, Univ. Cambridge (2012).
Chen, P.R. et al. Angew. Chem. Int. Edn. Engl. 48, 4052–4055 (2009).
Mukai, T. et al. Biochem. Biophys. Res. Commun. 371, 818–822 (2008).
Greiss, S. & Chin, J.W. J. Am. Chem. Soc. 133, 14196–14199 (2011).
Isogai, Y., Takada, S., Tjian, R. & Keles, S. EMBO J. 26, 79–89 (2007).
Mukai, T., Wakiyama, M., Sakamoto, K. & Yokoyama, S. Protein Sci. 19, 440–448 (2010).
Bunch, T.A., Grinblat, Y. & Goldstein, L.S. Nucleic Acids Res. 16, 1043–1061 (1988).
Van Doren, M., Williamson, A.L. & Lehmann, R. Curr. Biol. 8, 243–246 (1998).
Pignoni, F. & Zipursky, S.L. Development 124, 271–278 (1997).
Acknowledgements
This work was funded by the UK Medical Research Council (MRC) (grants U105181009 and UD99999908) and the Louis-Jeantet Foundation Young Investigator Career Award (both to J.W.C.). We are grateful to S. Bullock (MRC Laboratory of Molecular Biology) for providing initial plasmids and for extensive help and advice with this project; to S. Peak-Chew and M. Skehel in the MRC Laboratory of Molecular Biology Mass Spectrometry Facility for MS; S. Brunner for assistance with statistical analysis; and J. Schmitz and R. Pisa for experimental assistance.
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A.B., F.M.T. and S.G. designed and performed experiments. K.L. provided compounds. A.B. and J.W.C. wrote the paper with input from all authors.
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Bianco, A., Townsley, F., Greiss, S. et al. Expanding the genetic code of Drosophila melanogaster. Nat Chem Biol 8, 748–750 (2012). https://doi.org/10.1038/nchembio.1043
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DOI: https://doi.org/10.1038/nchembio.1043
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