biostudies-literatureStonkus JResearch Council of Lithuaniagkaf1379https://www.ebi.ac.uk/biostudies/studies/S-EPMC12709175biostudies-literatureUnknown53(22)Nucleoside 2'-O-methylation is a widespread RNA modification found across diverse RNA types in all domains of life. Although considerable progress has been made in mapping 2'-O-methylation sites and elucidating their biological roles, the enzymatic pathways responsible for the catabolism of 2'-O-methylated nucleosides remain largely unexplored. Here, we report a previously unidentified 2'-O-methylated nucleoside demethylation reaction catalyzed by a metagenome-derived bacterial dioxygenase named FJS. We further show that FJS-related enzymes, including human phytanoyl-CoA dioxygenase domain-containing protein 1 (PHYHD1), catalyze the 2-oxoglutarate-dependent demethylation of a broad range of ribose-methylated nucleosides, but do not act on modified nucleotides or 3'-terminal 2'-O-methylatedNucleic acids researchA new role for PHYHD1 and related dioxygenases: demethylation of 2'-O-methylated nucleosides.PMC12709175S-MIP-24-50Rutkiene RJasiuniene MKaliniene LLazutka JMeskys RAucynaite AVilkaitis GStonkus JBalciunas DMeskiene RfalseA new role for PHYHD1 and related dioxygenases: demethylation of 2'-O-methylated nucleosides.Nucleoside 2'-O-methylation is a widespread RNA modification found across diverse RNA types in all domains of life. Although considerable progress has been made in mapping 2'-O-methylation sites and elucidating their biological roles, the enzymatic pathways responsible for the catabolism of 2'-O-methylated nucleosides remain largely unexplored. Here, we report a previously unidentified 2'-O-methylated nucleoside demethylation reaction catalyzed by a metagenome-derived bacterial dioxygenase named FJS. We further show that FJS-related enzymes, including human phytanoyl-CoA dioxygenase domain-containing protein 1 (PHYHD1), catalyze the 2-oxoglutarate-dependent demethylation of a broad range of ribose-methylated nucleosides, but do not act on modified nucleotides or 3'-terminal 2'-O-methylated2025-01-01T00:00:00Z2025 Nov2026-06-08T03:19:57.943Z2026-06-08T03:09:15.586ZS-EPMC127091754140480110.1093/nar/gkaf1379