Discrimination among E. coli tRNAs with a long variable arm.

Abstract

In E. coli, tRNA(Ser), tRNA(Leu) and tRNA(Tyr) have a long variable arm composed of more than ten nucleotides (class II tRNAs). In order to study how leucyl- and seryl-tRNA synthetase discriminate their cognate tRNA isoacceptors from the other class II tRNAs, kinetic parameters of various mutated class II tRNA transcripts with leucyl- and seryl-tRNA synthetase were determined. Leucyl-tRNA synthetase recognizes A73 and A14 or its vicinity. Seryl-tRNA synthetase recognizes the long variable arm base-nonspecifically. C2-G71 in the acceptor stem functions as a negative identity element against seryl-tRNA synthetase. Difference in the tertiary structure among class II tRNA molecules plays a crucial role in discrimination by these two synthetases.