ZFIN Image: Stonkus et al., 2025, Figure 1.

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Figure Caption

Figure 1.

Discovery and characterization of FJS catalytic activity. (A) The functionality of FJS, encoded in the pUC19 vector, was identified using the E. coli DH10B ΔpyrFEC strain. (B) The enzymatic activity of FJS depends on 2-oxoglutarate and is further enhanced in vitro by adding ascorbate and FeSO₄. (C) HPLC-MS analysis confirmed the loss of the methyl group during the reaction by detecting a 14 Da decrease in the mass of 2′-O-methylcytidine. (D) The methyl group from the modified nucleoside is converted to formaldehyde, as demonstrated by the reaction with Purpald reagent. (E) Proposed reaction scheme for the FJS-catalyzed demethylation of 2′-O-methylcytidine. Cm, 2′-O-methylcytidine; C, cytidine; 2OG, 2-oxoglutarate; Asc, ascorbate.

Acknowledgments

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