Natural human tRNA^Ala anticodon variants mistranslate the genetic code

Tandem mass spectrometry-based estimation of mistranslation levels in cells with tRNA^Ala variants. The plots show the proportion of mistranslated peptides in mCherry from HEK 293T cells expressing the indicated wild-type or mistranslating tRNA^Ala variants. (A, B) The percentage of Ala misincorporation was calculated by comparing the area under the isotopic peak observed for matching mistranslated relative to properly translated peptides and presented in (A) standard and (B) logarithmic plots. Independent sample t-tests were (* P < 0.05, ** P < 0.01, *** P < 0.001) based on at least n = 3 biological replicates.