When the primitive translation system first emerged in the hypothetical RNA world, ribozymes could have been responsible for aminoacylation. Given that naturally occurring T-box riboswitches selectively sense the aminoacylation status of cognate tRNAs, we introduced a domain of random sequence into a T-box–tRNA conjugate and isolated ribozymes that were self-aminoacylating on the 3′-terminal hydroxyl group. One of them, named Tx2.1, recognizes the anticodon and D-loop of tRNA via interaction with its stem I domain, similarly to the parental T-box, and selectively charges N-biotinyl-l-phenylalanine (Bio-lPhe) onto the 3′ end of the cognate tRNA in trans. We also demonstrated the ribosomal synthesis of a Bio-lPhe-initiated peptide in a Tx2.1-coupled in vitro translation system, in which Tx2.1 catalyzed specific tRNA aminoacylation in situ. This suggests that such ribozymes could have coevolved with a primitive translation system in the RNA world.
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This work was supported by Japan Society for the Promotion of Science (JSPS) Grants-in-Aid for Research S (26220204), the Human Frontier Science Program (RGP0015/2017), the Japan Science and Technology (JST) agency and CREST-Molecular Technology (JPMJCR12L2) to H.S. N.T. is supported by Grants-in-Aid for JSPS Fellows (12J08188) and Grants-in-Aid for Early-Career Scientists (19K16200). S.I. was supported by Grants-in-Aid for JSPS Fellows (JP16J04031). T.K. is supported by a JSPS Grant-in-Aid for Challenging Exploratory Research (26560429).
The authors declare no competing interests.
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Ishida, S., Terasaka, N., Katoh, T. et al. An aminoacylation ribozyme evolved from a natural tRNA-sensing T-box riboswitch. Nat Chem Biol 16, 702–709 (2020). https://doi.org/10.1038/s41589-020-0500-6
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