TRNA anticodons with the modified nucleoside 2-methylthio- N6-(4-hydroxyisopentenyl)adenosine distinguish between bases 3′ of the codon

Abstract

The modified nucleoside 2-methylthio- N 6-(4-hydroxyisopentenyl)adenosine (ms 2io 6A) is present immediately to the 3′ side of the anticodon (position 37) in tRNAs that read codons starting with uridine and hence include amber (UAG) suppressor tRNAs. We have used strains of Salmonella typhimurium that differ only in their ability to synthesize ms 2io 6A in order to determine specifically how this modified nucleoside influences the efficiency of amber suppression in two codon contexts differing by only which base is 3′ of the codon. The results show that the presence of the modified nucleoside ms 2io 6A not only improves the efficiency of the suppressor tRNAs but also allows them to distinguish between at least two bases 3′ of the codon. Thus, the presence of ms 2io 6A reduces the intrinsic codon context sensitivity of the tRNA and specifically counteracts an unfavourable nucleotide on the 3′ side of the codon. The possible codon-anticodon interactions responsible for this effect are discussed.