Chemical Method to Sequence 5-Formylcytosine on RNA.

TitleChemical Method to Sequence 5-Formylcytosine on RNA.
Publication TypeJournal Article
Year of Publication2022
AuthorsLi, A, Sun, X, A Arguello, E, Kleiner, RE
JournalACS Chem Biol
Volume17
Issue3
Pagination503-508
Date Published2022 Mar 18
ISSN1554-8937
KeywordsAnimals, Cytosine, Mammals, RNA, RNA Processing, Post-Transcriptional, RNA, Transfer
Abstract

<p>Epitranscriptomic RNA modifications can regulate biological processes, but there remains a major gap in our ability to identify and measure individual modifications at nucleotide resolution. Here we present Mal-Seq, a chemical method for sequencing 5-formylcytosine (fC) modifications on RNA based on the selective and efficient malononitrile-mediated labeling of fC residues to generate adducts that are read as C-to-T mutations upon reverse transcription and polymerase chain reaction amplification. We apply Mal-Seq to characterize the prevalence of fC at the wobble position of mt-tRNA(Met) in different organisms and tissue types and find that high-level fC modification is present in mammals but lacking in lower eukaryotes. Our work sheds light on mitochondrial tRNA modifications throughout eukaryotic evolution and provides a general platform for characterizing the fC epitranscriptome.</p>

DOI10.1021/acschembio.1c00707
Alternate JournalACS Chem Biol
PubMed ID35212224
PubMed Central IDPMC9357364
Grant ListR01 GM132189 / GM / NIGMS NIH HHS / United States