MassIVE MSV000091065

Partial Public PXD039448

Environment Modulates Protein Heterogeneity Through Transcriptional and Translational Stop Codon Recoding

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Description

Stop codon recoding events give rise to longer proteins, which may alter the proteins function and thereby generate short-lasting phenotypic variability from a single gene. In order to systematically assess the frequency and origin of recoding events, we designed a library of reporters. We introduced premature stop codons into mScarlet that enabled high-throughput quantification of protein synthesis termination errors in E.coli using fluorescent microscopy. We found that under stress conditions, stop codon recoding may occur as high as 80 percent of the time, depending on the genetic context, suggesting that evolution frequently samples stop codon recoding events. Targeted mass spectrometry and RNA-seq analyses showed that not only translational but also transcriptional errors contribute to stop codon recoding. The RNA polymerase is more likely to misincorporate a nucleotide at premature stop codons. Proteome-wide mass -spectrometry revealed that temperature regulates the expression of cryptic peptides generated by stop codon recoding in E.coli. Overall, our findings suggest that the environment influences the accuracy of protein production which increases protein heterogeneity when the organisms need to adapt to new conditions. [doi:10.25345/C5TM72991] [dataset license: CC0 1.0 Universal (CC0 1.0)]

Keywords: phenotypic mutations ; stop codon readthrough ; RNA polymerase error

Contact

Principal Investigators:
(in alphabetical order) 		Agnes Toth-Petroczy, MPI-CBG, Germany
Submitting User: Tobias
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