Environment modulates protein heterogeneity through transcriptional and translational stop codon readthrough.
Journal
Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555
Informations de publication
Date de publication:
24 May 2024
24 May 2024
Historique:
received:
22
02
2023
accepted:
25
04
2024
medline:
25
5
2024
pubmed:
25
5
2024
entrez:
24
5
2024
Statut:
epublish
Résumé
Stop codon readthrough events give rise to longer proteins, which may alter the protein's function, thereby generating short-lasting phenotypic variability from a single gene. In order to systematically assess the frequency and origin of stop codon readthrough events, we designed a library of reporters. We introduced premature stop codons into mScarlet, which enabled high-throughput quantification of protein synthesis termination errors in E. coli using fluorescent microscopy. We found that under stress conditions, stop codon readthrough may occur at rates as high as 80%, depending on the nucleotide context, suggesting that evolution frequently samples stop codon readthrough events. The analysis of selected reporters by mass spectrometry and RNA-seq showed that not only translation but also transcription errors contribute to stop codon readthrough. The RNA polymerase was more likely to misincorporate a nucleotide at premature stop codons. Proteome-wide detection of stop codon readthrough by mass spectrometry revealed that temperature regulated the expression of cryptic sequences generated by stop codon readthrough in E. coli. Overall, our findings suggest that the environment affects the accuracy of protein production, which increases protein heterogeneity when the organisms need to adapt to new conditions.
Identifiants
pubmed: 38789441
doi: 10.1038/s41467-024-48387-x
pii: 10.1038/s41467-024-48387-x
doi:
Substances chimiques
Codon, Terminator
0
Escherichia coli Proteins
0
Codon, Nonsense
0
DNA-Directed RNA Polymerases
EC 2.7.7.6
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
4446Informations de copyright
© 2024. The Author(s).
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