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Ochkasova, Anastasia S.; Meschaninova, Maria I.; Venyaminova, Aliya G.; Graifer, Dmitri M.; Karpova, Galina G.
Biochimica et biophysica acta. Proteins and proteomics, October 2021, 2021-10-00, 20211001, Letnik: 1869, Številka: 10Journal Article
Abasic (AP) sites in mRNAs are lesions whose accumulation in cells is linked to various neurodegenerative diseases arising from the appearance of truncated peptides due to the premature cessation of translation of these mRNAs. It is believed that the translation of AP site-containing mRNAs is stopped when the damaged codon arrives to the A site, where it is not decoded. We propose an alternative translation arrest mechanism mediated by the 40S ribosomal subunit protein uS3. Recently, it has been shown that in human 80S ribosomal complexes assembled without translation factors, uS3 cross-links to the AP site at the 3′-terminus of the mRNA, whose undamaged part is bound at the 40S subunit channel, via its peptide 55–64 exposed near the mRNA entry pore. In this study, we examined whether such cross-linking occurs during the translation of mRNA with the AP site. To this end, we used a set of synthetic mRNAs bearing the AP site inserted in the desired location in their sequences. An analysis of 80S ribosomal complexes formed with these mRNAs in a mammalian cell-free protein-synthesizing system demonstrates that AP sites do indeed cross-link to uS3 in the course of the translation. We also show that the cross-linking occurs as soon as the AP site arrives to a common favorable position relative to uS3, which is independent on its location in the mRNA. Our findings suggest that the mechanism of stopping translation of damaged mRNAs involving uS3, along with the one mentioned above, could underlie ribosome-associated mRNA quality control. •Damaged mRNA cross-links to ribosomal protein uS3 via abasic site during translation.•mRNA abasic site cross-links to uS3 when arrives to the certain favorable position.•The 55–64 peptide of uS3 inspects mRNAs for the presence of abasic sites during translation.•Abasic site-uS3 cross-linking is one of ways of the ribosome-based mRNA surveillance.
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