Designer ribozymes: programming the tRNA specificity into flexizyme |
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Authors: | Ramaswamy Krishna Saito Hirohide Murakami Hiroshi Shiba Kiyotaka Suga Hiroaki |
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Affiliation: | Departments of Chemistry and Biological Sciences, University at Buffalo, State University of New York, Buffalo, NY 14260-3000, USA. |
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Abstract: | Fx3 is an artificial ribozyme with the ability to aminoacylate various tRNAs with phenylalanine and its nonnatural derivatives. Herein we report a simple strategy to build tRNA specificity into the generic Fx3, by appending to its 3'-end a tRNA-specific sequence (TSS), which is complementary to the acceptor stem of the cognate tRNA. This new designer ribozyme, referred to as Fx10, is able to recognize its cognate tRNA via a 10-base-pair interaction that is formed after the invasion of the tRNA acceptor stem by the TSS. We have demonstrated that Fx10 can aminoacylate its cognate tRNA with a high degree of specificity and also discriminate against the noncognate tRNAs. Because the tRNA specificity can be easily programmed into Fx10, it is a custom-made catalyst to generate nonnatural aminoacyl-tRNAs. |
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