Designer ribozymes: programming the tRNA specificity into flexizyme.
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.
Ramaswamy
Krishna
K
Departments of Chemistry and Biological Sciences, University at Buffalo, State University of New York, Buffalo, NY 14260-3000, USA.
Saito
Hirohide
H
Murakami
Hiroshi
H
Shiba
Kiyotaka
K
Suga
Hiroaki
H
D000215:Acylation; D005818:Genetic Engineering / Q000379:methods; D009690:Nucleic Acid Conformation; D016337:RNA, Catalytic / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D012343:RNA, Transfer / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity
D016337:RNA, Catalytic; D012343:RNA, Transfer