Title | Chemical Method to Sequence 5-Formylcytosine on RNA. |
Publication Type | Journal Article |
Year of Publication | 2022 |
Authors | Li, A, Sun, X, A Arguello, E, Kleiner, RE |
Journal | ACS Chem Biol |
Volume | 17 |
Issue | 3 |
Pagination | 503-508 |
Date Published | 2022 03 18 |
ISSN | 1554-8937 |
Keywords | Animals, 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> |
DOI | 10.1021/acschembio.1c00707 |
Alternate Journal | ACS Chem Biol |
PubMed ID | 35212224 |
Grant List | R01 GM132189 / GM / NIGMS NIH HHS / United States |