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Structural basis for orthogonal tRNA specificities of tyrosyl-tRNA synthetases for genetic code expansion
Abstract
The archaeal/eukaryotic tyrosyl-tRNA synthetase (TyrRS)–tRNATyr pairs do not cross-react with their bacterial counterparts. This 'orthogonal' condition is essential for using the archaeal pair to expand the bacterial genetic code. In this study, the structure of the Methanococcus jannaschii TyrRS–tRNATyr–L-tyrosine complex, solved at a resolution of 1.95 Å, reveals that this archaeal TyrRS strictly recognizes the C1-G72 base pair, whereas the bacterial TyrRS recognizes the G1-C72 in a different manner using different residues. These diverse tRNA recognition modes form the basis for the orthogonality. The common tRNATyr identity determinants (the discriminator, A73 and the anticodon residues) are also recognized in manners different from those of the bacterial TyrRS. Based on this finding, we created a mutant TyrRS that aminoacylates the amber suppressor tRNA with C34 65 times more efficiently than does the wild-type enzyme.
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Cites
Universal rules and idiosyncratic features in tRNA identity
Giege, R., Sissler, M., & Florentz, C. (1998) · Nucleic Acids Research · doi:10.1093/nar/26.22.5017 · Cited by 769
Lu, W., Gong, D., Bar-Sagi, D., & Cole, P. A. (2001) · Molecular Cell · doi:10.1016/s1097-2765(01)00369-0 · Cited by 268
Cited by
Adding New Chemistries to the Genetic Code
Liu, C. C., & Schultz, P. G. (2010) · Annual Review of Biochemistry · doi:10.1146/annurev.biochem.052308.105824 · Cited by 1,725
A chemical toolkit for proteins — an expanded genetic code
Xie, J., & Schultz, P. G. (2006) · Nature Reviews Molecular Cell Biology · doi:10.1038/nrm2005 · Cited by 408
Metadata
- Authors
- Kobayashi, T., Nureki, O., Ishitani, R., Yaremchuk, A., Tukalo, M., Cusack, S., Sakamoto, K., & Yokoyama, S.
- Venue
- Nature Structural & Molecular Biology
- Published
- 2003-05-19
- Volume
- 10
- Issue
- 6
- Pages
- 425-432
- ISSN
- 1545-9985 (ISSN-L)
- 1545-9993
- OpenAlex
- W2061539493
- Cited by
- 204
- References
- 2
- FWCI
- 4.61
- Type
- journal article
- Language
- en
- Open access
- closed
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- Kobayashi, T., Nureki, O., Ishitani, R., Yaremchuk, A., Tukalo, M., Cusack, S., Sakamoto, K., & Yokoyama, S. (2003). Structural basis for orthogonal tRNA specificities of tyrosyl-tRNA synthetases for genetic code expansion. Nature Structural & Molecular Biology. https://doi.org/10.1038/nsb934
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