Seryl-tRNA synthetase promotes translational readthrough by mRNA binding and involvement of the selenocysteine incorporation machinery

Translational readthrough of UGA stop codons by selenocysteine-specific tRNA (tRNASec) enables the synthesis of selenoproteins. Seryl-tRNA synthetase (SerRS) charges tRNASec with serine, which is modified into selenocysteine and delivered to the ribosome by a designated elongation factor (eEFSec in eukaryotes). Here we found that components of the human selenocysteine incorporation machinery (SerRS, tRNASec, and eEFSec) also increased translational readthrough of non-selenocysteine genes, including VEGFA, to create C-terminally extended isoforms. SerRS recognizes target mRNAs through a stem-loop structure that resembles the variable loop of its cognate tRNAs. This function of SerRS depends on both its enzymatic activity and a vertebrate-specific domain. Through eCLIP-seq, we identified additional SerRS-interacting mRNAs as potential readthrough genes. Moreover, SerRS overexpression was sufficient to reverse premature termination caused by a pathogenic nonsense mutation. Our findings expand the repertoire of selenoprotein biosynthesis machinery and suggest an avenue for therapeutic targeting of nonsense mutations using endogenous factors. Graphical Abstract

Automated summary

Translational readthrough of UGA stop codons by selenocysteine-specific tRNA enables the synthesis of selenoproteins. In addition, this mechanism can also increase translational readthrough of non-selenocysteine genes to create C-terminally extended isoforms. The recognition of target mRNAs by seryl-tRNA synthetase is dependent on its enzymatic activity and a vertebrate-specific domain. The findings expand our understanding of selenoprotein biosynthesis and suggest a potential avenue for therapeutic targeting of nonsense mutations using endogenous factors.