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      Identification and quantification of modified nucleosides in Saccharomyces cerevisiae mRNAs

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      ACS Chemical Biology

      American Chemical Society (ACS)

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          Abstract

          Post-transcriptional modifications to messenger RNAs (mRNAs) have the potential to alter the biological function of this important class of biomolecules. The study of mRNA modifications is a rapidly emerging field, and the full complement of chemical modifications in mRNAs is not yet established. We sought to identify and quantify the modifications present in yeast mRNAs using an ultra-high performance liquid chromatography tandem mass spectrometry method to detect 40 nucleoside variations in parallel. We observe six modified nucleosides with high confidence in highly purified mRNA samples (N7-methylguanosine, N6-methyladenosine, 2′- O -methylguanosine, 2′- O -methylcytidine, N4-acetylcytidine, and 5-formylcytidine) and identify the yeast protein responsible for N4-acetylcytidine incorporation in mRNAs (Rra1). In addition, we find that mRNA modification levels change in response to heat shock, glucose starvation, and/or oxidative stress. This work expands the repertoire of potential chemical modifications in mRNAs and highlights the value of integrating mass spectrometry tools in the mRNA modification discovery and characterization pipeline.

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          Author and article information

          Journal
          ACS Chemical Biology
          ACS Chem. Biol.
          American Chemical Society (ACS)
          1554-8929
          1554-8937
          June 19 2019
          June 19 2019
          Article
          10.1021/acschembio.9b00369
          7254066
          31243956
          bbaa0d00-cdd1-4d95-8dc4-55d72134c300
          © 2019
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