Meclofenamic acid selectively inhibits FTO demethylation of m6A over ALKBH5
Open Access
- 1 December 2014
- journal article
- research article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 43 (1), 373-384
- https://doi.org/10.1093/nar/gku1276
Abstract
Two human demethylases, the fat mass and obesity-associated (FTO) enzyme and ALKBH5, oxidatively demethylate abundant N6-methyladenosine (m6A) residues in mRNA. Achieving a method for selective inhibition of FTO over ALKBH5 remains a challenge, however. Here, we have identified meclofenamic acid (MA) as a highly selective inhibitor of FTO. MA is a non-steroidal, anti-inflammatory drug that mechanistic studies indicate competes with FTO binding for the m6A-containing nucleic acid. The structure of FTO/MA has revealed much about the inhibitory function of FTO. Our newfound understanding, revealed herein, of the part of the nucleotide recognition lid (NRL) in FTO, for example, has helped elucidate the principles behind the selectivity of FTO over ALKBH5. Treatment of HeLa cells with the ethyl ester form of MA (MA2) has led to elevated levels of m6A modification in mRNA. Our collective results highlight the development of functional probes of the FTO enzyme that will (i) enable future biological studies and (ii) pave the way for the rational design of potent and specific inhibitors of FTO for use in medicine.Keywords
This publication has 64 references indexed in Scilit:
- ALKBH5 Is a Mammalian RNA Demethylase that Impacts RNA Metabolism and Mouse FertilityMolecular Cell, 2013
- Comprehensive Analysis of mRNA Methylation Reveals Enrichment in 3′ UTRs and near Stop CodonsCell, 2012
- Duplex interrogation by a direct DNA repair protein in search of base damageNature Structural & Molecular Biology, 2012
- Exocyclic Carbons Adjacent to the N6 of Adenine are Targets for Oxidation by the Escherichia coli Adaptive Response Protein AlkBJournal of the American Chemical Society, 2012
- N6-Methyladenosine in nuclear RNA is a major substrate of the obesity-associated FTONature Chemical Biology, 2011
- A Selective Inhibitor and Probe of the Cellular Functions of Jumonji C Domain-Containing Histone DemethylasesJournal of the American Chemical Society, 2011
- The AlkB Domain of Mammalian ABH8 Catalyzes Hydroxylation of 5‐Methoxycarbonylmethyluridine at the Wobble Position of tRNAAngewandte Chemie-International Edition, 2010
- Oxidative demethylation of 3‐methylthymine and 3‐methyluracil in single‐stranded DNA and RNA by mouse and human FTOFEBS Letters, 2008
- Crystal structures of DNA/RNA repair enzymes AlkB and ABH2 bound to dsDNANature, 2008
- [20] Processing of X-ray diffraction data collected in oscillation modeMethods in Enzymology, 1997