Quality control of protein synthesis in the early elongation stage

Asuteka Nagao (), Yui Nakanishi, Yutaro Yamaguchi, Yoshifumi Mishina, Minami Karoji, Takafumi Toya, Tomoya Fujita, Shintaro Iwasaki, Kenjyo Miyauchi, Yuriko Sakaguchi and Tsutomu Suzuki ()
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Asuteka Nagao: University of Tokyo
Yui Nakanishi: University of Tokyo
Yutaro Yamaguchi: University of Tokyo
Yoshifumi Mishina: University of Tokyo
Minami Karoji: University of Tokyo
Takafumi Toya: University of Tokyo
Tomoya Fujita: RIKEN Cluster for Pioneering Research
Shintaro Iwasaki: RIKEN Cluster for Pioneering Research
Kenjyo Miyauchi: University of Tokyo
Yuriko Sakaguchi: University of Tokyo
Tsutomu Suzuki: University of Tokyo

Nature Communications, 2023, vol. 14, issue 1, 1-20

Abstract: Abstract In the early stage of bacterial translation, peptidyl-tRNAs frequently dissociate from the ribosome (pep-tRNA drop-off) and are recycled by peptidyl-tRNA hydrolase. Here, we establish a highly sensitive method for profiling of pep-tRNAs using mass spectrometry, and successfully detect a large number of nascent peptides from pep-tRNAs accumulated in Escherichia coli pthts strain. Based on molecular mass analysis, we found about 20% of the peptides bear single amino-acid substitutions of the N-terminal sequences of E. coli ORFs. Detailed analysis of individual pep-tRNAs and reporter assay revealed that most of the substitutions take place at the C-terminal drop-off site and that the miscoded pep-tRNAs rarely participate in the next round of elongation but dissociate from the ribosome. These findings suggest that pep-tRNA drop-off is an active mechanism by which the ribosome rejects miscoded pep-tRNAs in the early elongation, thereby contributing to quality control of protein synthesis after peptide bond formation.

Date: 2023
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DOI: 10.1038/s41467-023-38077-5

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