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bioRxiv · 10.1101/2022.04.27.489667

Novel trans-translation-associated gene regulation revealed by prophage excision-triggered switching of ribosome rescue pathway

Onodera, H.·Niwa, T.·Taguchi, H.·Chadani, Y.
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Abstract

Escherichia coli has multiple pathways to release a nonproductive ribosome complex stalled at the 3 end of nonstop mRNAs: SsrA RNA-mediated trans-translation and stop codon-independent termination by ArfA/RF2 or ArfB (YaeJ). The arfA mRNA lacks a stop codon and thus its expression is repressed by trans-translation. Therefore, ArfA is considered to complement the ribosome rescue activity of trans-translation, but the situations in which ArfA is expressed to rescue the nonproductive complexes have not been elucidated. Here, we demonstrated that the excision of the CP4-57 prophage adjacent to the E. coli ssrA gene leads to the inactivation of SsrA RNA and switches the primary rescue pathway from trans-translation to the ArfA/RF2 pathway. A comparative quantitative proteomic analysis revealed that the switching of the rescue pathway rearranges not only the proteome landscape in E. coli cells but also the phenotype, such as motility. Among the proteins with significantly increased abundance in the ssrA-inactivated cells, we found ZntR, whose mRNA is transcribed together as the upstream part of the nonstop arfA mRNA. Further analysis revealed that the translation of the nonstop ORF of arfA triggered the repression of the upstream zntR ORF, via trans-translation-coupled exonucleolytic (RUTEX) degradation by a polynucleotide phosphorylase. These results provide a novel example of trans-translation-dependent regulation, and shed new light on the physiological roles of prophages in gene expression.

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Onodera, H., Niwa, T., Taguchi, H., Chadani, Y.. 2022-04-28. Novel trans-translation-associated gene regulation revealed by prophage excision-triggered switching of ribosome rescue pathway. https://doi.org/10.1101/2022.04.27.489667

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