Abstract
N6-methyladenosine (m6A) is a prevalent internal modification in mRNA and non- coding RNA affecting various cellular pathways. Here we report the discovery of two additional modifications, N6-hydroxymethyladenosine (hm6A) and N6- formyladenosine (f6A), in mammalian mRNA. We show that FeII- and α-ketoglutarate (α-KG)-dependent fat mass and obesity associated (FTO) protein oxidizes m6A to generates hm6A as an intermediate modification and f6A as a further oxidized product. hm6A and f6A have half-life times of ~3 h in aqueous solution under physiological relevant conditions, and are present in isolated mRNA from human cells as well as mouse tissues. These previously unknown modifications derived from the prevalent m6A in mRNA, formed through oxidative RNA demethylation, may dynamically modulate RNA-protein interactions to affect gene expression regulation.
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