FTO-mediated formation of N6-hydroxymethyladenosine and N6-formyladenosine in mammalian RNA

Nat Commun. 2013:4:1798. doi: 10.1038/ncomms2822.


N(6)-methyladenosine is a prevalent internal modification in messenger RNA and non-coding RNA affecting various cellular pathways. Here we report the discovery of two additional modifications, N(6)-hydroxymethyladenosine (hm(6)A) and N(6)-formyladenosine (f(6)A), in mammalian messenger RNA. We show that Fe(II)- and α-ketoglutarate-dependent fat mass and obesity-associated (FTO) protein oxidize N(6)-methyladenosine to generate N(6)-hydroxymethyladenosine as an intermediate modification, and N(6)-formyladenosine as a further oxidized product. N(6)-hydroxymethyladenosine and N(6)-formyladenosine have half-life times of ~3 h in aqueous solution under physiological relevant conditions, and are present in isolated messenger RNA from human cells as well as mouse tissues. These previously unknown modifications derived from the prevalent N(6)-methyladenosine in messenger RNA, formed through oxidative RNA demethylation, may dynamically modulate RNA-protein interactions to affect gene expression regulation.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Adenosine / analogs & derivatives*
  • Adenosine / chemistry
  • Adenosine / metabolism
  • Alpha-Ketoglutarate-Dependent Dioxygenase FTO
  • Animals
  • Humans
  • Kinetics
  • Mammals / metabolism*
  • Methylation
  • Mice
  • Models, Biological
  • Molecular Dynamics Simulation
  • Oxidation-Reduction
  • Protein Binding
  • Proteins / metabolism*
  • RNA / chemistry
  • RNA / metabolism*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • RNA-Binding Proteins / metabolism
  • Substrate Specificity


  • Proteins
  • RNA, Messenger
  • RNA-Binding Proteins
  • RNA
  • N(6)-hydroxymethyladenosine
  • Alpha-Ketoglutarate-Dependent Dioxygenase FTO
  • FTO protein, human
  • Adenosine