Misacylation of tRNA in prokaryotes: a re-evaluation

Abstract

Misacylation of tRNA by a non-cognate amino acid is a natural phenomenon and occurs with a frequency of approximately 1 in 10,000 due to occasional mistakes in aminoacyl transfer RNA (tRNA) synthesis. In a number of prokaryotic organisms, misacylation of selenocysteinyl tRNA, glutaminyl tRNA and aspartyl tRNAs has particular physiological meaning. Recently, misacylation has emerged as a powerful tool for studying specific interactions between aa-tRNAs and associated protein factors. The present review provides an overview of the application of misacylated tRNA in research.

Cotranslational incorporation of non‐standard amino acids using cell‐free protein synthesis

“…The term “misacylation” refers to the aminoacylation of a tRNA with a non‐cognate amino acid [6,56]. In their natural context, tRNAs are enzymatically aminoacylated with their cognate amino acids by highly specific aminoacyl‐tRNA synthetases (aaRS) [57].…”

Class I and class II lysyl‐tRNA synthetase mutants and the genetic encoding of pyrrolysine in Methanosarcina spp.

“…Recent papers have commented on the presence of both charging systems for tRNA Pyl within Methanosarcina spp. (Namy et al ., 2004; Veit et al ., 2005; Klotz et al ., 2006; Stortchevoi, 2006; Delarue, 2007), and the need to evaluate the in vivo significance of LysRS1/LysRS2 charging of tRNA Pyl (Stortchevoi, 2006).…”

Synchronized Translation for Detection of Temporal Stalling of Ribosome during Single-Turnover Translation

“…Under the condition, the ribosome might misincorporate an amino acid other than isoleucine at the thirty-second AUC condon via a near-cognate aminoacyl-tRNA or tRNA Ile misaminoacylated by another amionacyl-tRNA synthetase. 12 When the tRNA concentration was diluted, the signal of the full-length product became weaker and a fluorescence signal with lower molecular weight (Figure 2a, H) was observed. The position corresponded to a peptide mass of the expected product, in which the ribosome halted before the isoleucine codon, in that case, translated peptide containing 31 amino acids should have a mass of 4064.4 Da.…”

“…The position was the same as the translated product performed in the PURE system containing all amionacyl-tRNA synthetases (Figure S-1, Supporting Information), indicating that this product resulted from translation through the 3′ end of the mRNA. Under the condition, the ribosome might misincorporate an amino acid other than isoleucine at the thirty-second AUC condon via a near-cognate aminoacyl-tRNA or tRNA Ile misaminoacylated by another amionacyl-tRNA synthetase . When the tRNA concentration was diluted, the signal of the full-length product became weaker and a fluorescence signal with lower molecular weight (Figure a, H) was observed.…”