Designer ribozymes: programming the tRNA specificity into flexizyme

Krishna Ramaswamy; Hirohide Saito; Hiroshi Murakami; Kiyotaka Shiba; Hiroaki Suga

doi:10.1021/ja046843y

Designer ribozymes: programming the tRNA specificity into flexizyme

J Am Chem Soc. 2004 Sep 22;126(37):11454-5. doi: 10.1021/ja046843y.

Authors

Krishna Ramaswamy¹, Hirohide Saito, Hiroshi Murakami, Kiyotaka Shiba, Hiroaki Suga

Affiliation

¹ Departments of Chemistry and Biological Sciences, University at Buffalo, State University of New York, Buffalo, NY 14260-3000, USA.

PMID: 15366888
DOI: 10.1021/ja046843y

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Acylation
Genetic Engineering / methods
Nucleic Acid Conformation
RNA, Catalytic / chemistry*
RNA, Catalytic / genetics
RNA, Catalytic / metabolism
RNA, Transfer / chemistry*
RNA, Transfer / genetics
RNA, Transfer / metabolism
Substrate Specificity

Substances

RNA, Catalytic
RNA, Transfer