YTHDF2 Recognition of N-Methyladenosine (mA)-Modified RNA Is Associated with Transcript Destabilization.

TitleYTHDF2 Recognition of N-Methyladenosine (mA)-Modified RNA Is Associated with Transcript Destabilization.
Publication TypeJournal Article
Year of Publication2020
AuthorsSeo, KW, Kleiner, RE
JournalACS Chem Biol
Date Published2020 Jan 17
KeywordsAdenosine, Amino Acid Sequence, Gene Expression Regulation, Methylation, Protein Binding, Protein Biosynthesis, Protein Interaction Domains and Motifs, Recombinant Proteins, RNA Stability, RNA, Messenger, RNA-Binding Proteins, Transcription Factors

<p>Epitranscriptomic modifications play an important role in RNA function and can impact gene expression. Here, we apply a chemical proteomics approach to investigate readers of N-methyladenosine (mA), a poorly characterized modification on mammalian mRNA. We find that YTHDF proteins, known mA readers, recognize mA-modified sequences in a methylation-specific manner. We characterize binding of recombinant YTHDF1/2 proteins to mA-modified oligonucleotides to demonstrate that these interactions can exhibit comparable affinity to mA-recognition events and occur in diverse sequence contexts. Further, we demonstrate YTHDF2 interacts specifically with endogenously modified mA transcripts. Finally, we deplete cellular YTHDF2 to show that the abundance of mA-modified transcripts is increased in its absence. Similarly, increasing mA levels through depletion of ALKBH3, an mA eraser protein, destabilizes known mA-containing RNAs. Our results shed light on the function of mA on mRNA and provide a mechanistic framework to further evaluate the role of mA in biological processes.</p>

Alternate JournalACS Chem Biol
PubMed ID31815430
PubMed Central IDPMC7025767
Grant ListR01 GM132189 / GM / NIGMS NIH HHS / United States