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- W3028746489 endingPage "e71" @default.
- W3028746489 startingPage "e71" @default.
- W3028746489 abstract "Abstract The dynamic conformation of RNA molecules within living cells is key to their function. Recent advances in probing the RNA structurome in vivo, including the use of SHAPE (Selective 2′-Hydroxyl Acylation analyzed by Primer Extension) or kethoxal reagents or DMS (dimethyl sulfate), provided unprecedented insights into the architecture of RNA molecules in the living cell. Here, we report the establishment of lead probing in a global RNA structuromics approach. In order to elucidate the transcriptome-wide RNA landscape in the enteric pathogen Yersinia pseudotuberculosis, we combined lead(II) acetate-mediated cleavage of single-stranded RNA regions with high-throughput sequencing. This new approach, termed ‘Lead-seq’, provides structural information independent of base identity. We show that the method recapitulates secondary structures of tRNAs, RNase P RNA, tmRNA, 16S rRNA and the rpsT 5′-untranslated region, and that it reveals global structural features of mRNAs. The application of Lead-seq to Y. pseudotuberculosis cells grown at two different temperatures unveiled the first temperature-responsive in vivo RNA structurome of a bacterial pathogen. The translation of candidate genes derived from this approach was confirmed to be temperature regulated. Overall, this study establishes Lead-seq as complementary approach to interrogate intracellular RNA structures on a global scale." @default.
- W3028746489 created "2020-06-05" @default.
- W3028746489 creator A5045830997 @default.
- W3028746489 creator A5053881417 @default.
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- W3028746489 creator A5062030904 @default.
- W3028746489 creator A5078759067 @default.
- W3028746489 creator A5079185955 @default.
- W3028746489 creator A5082348781 @default.
- W3028746489 date "2020-05-28" @default.
- W3028746489 modified "2023-10-16" @default.
- W3028746489 title "Lead-seq: transcriptome-wide structure probing in vivo using lead(II) ions" @default.
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