A versatile tRNA aminoacylation catalyst based on RNA.
Aminoacyl-tRNA synthetase (ARS) ribozymes have potential to develop a novel genetic coding system. Although we have previously isolated such a ribozyme that recognizes aromatic amino acids, it could not be used as a versatile catalyst due to its limited ability of aminoacylation to a particular tRNA used for the selection. To overcome this limitation, we used a combination of evolutionary and engineering approaches to generate an optimized ribozyme. The ribozyme, consisting of 45 nucleotides, displays a broad spectrum of activity toward various tRNAs. Most significantly, this ribozyme is able to exhibit multiple turnover activity and charge parasubstituted Phe analogs onto an engineered suppressor tRNA (tRNA(Asn)(CCCG)). Thus, it provides a useful and flexible tool for the custom synthesis of mischarged tRNAs with natural and nonnatural amino acids.
Murakami
Hiroshi
H
Department of Chemistry, State University of New York, Buffalo, Buffalo, NY 14260, USA.
Saito
Hirohide
H
Suga
Hiroaki
H
D000596:Amino Acids / Q000737:chemistry*; D000604:Amino Acyl-tRNA Synthetases / Q000378:metabolism*; D002384:Catalysis; D002474:Cell-Free System; D003001:Cloning, Molecular; D007700:Kinetics; D009690:Nucleic Acid Conformation; D010649:Phenylalanine / Q000737:chemistry; D012313:RNA / Q000737:chemistry*; D016337:RNA, Catalytic / Q000378:metabolism*; D012343:RNA, Transfer / Q000737:chemistry*; D013329:Structure-Activity Relationship
D000596:Amino Acids; D016337:RNA, Catalytic; D010649:Phenylalanine; D012313:RNA; D012343:RNA, Transfer; D000604:Amino Acyl-tRNA Synthetases