SFPQ Is an FTO-Binding Protein that Facilitates the Demethylation Substrate Preference

The fat mass and obesity-associated protein (FTO) is the first identified demethylase of the internal RNA modification N⁶-methyladenosine (m⁶A), which also exhibits demethylation activity toward N⁶,2'-O-dimethyladenosine (m⁶A_m) and N¹-methyladenosine (m¹A). Demethylation of m⁶A at specific sites on target transcripts is a key enzymatic function of FTO that modulates diverse physiological and/or pathological processes. However, how FTO selects target RNA and whether additional interaction proteins facilitate this process remain elusive. Herein, via the genetically encoded and site-specific photocrosslinking strategy, we identified the major RNA-binding protein SFPQ as a direct interaction partner of FTO. Our study showed that FTO and SFPQ were located in close proximity throughout the transcriptome and that overexpression of SFPQ led to the demethylation of adjacent m⁶As, likely through recruiting FTO to these specific RNA sites. These results uncovered a new layer of regulation mechanism that may assist FTO to gain substrate specificity.