Activity-based RNA-modifying enzyme probing reveals DUS3L-mediated dihydrouridylation.

ABSTRACT: Epitranscriptomic RNA modifications can regulate RNA activity; however, there remains a major gap in our understanding of the RNA chemistry present in biological systems. Here we develop RNA-mediated activity-based protein profiling (RNABPP), a chemoproteomic strategy that relies on metabolic RNA labeling, mRNA interactome capture and quantitative proteomics, to investigate RNA-modifying enzymes in human cells. RNABPP with 5-fluoropyrimidines allowed us to profile 5-methylcytidine (m⁵C) and 5-methyluridine (m⁵U) methyltransferases. Further, we uncover a new mechanism-based crosslink between 5-fluorouridine (5-FUrd)-modified RNA and the dihydrouridine synthase (DUS) homolog DUS3L. We investigate the mechanism of crosslinking and use quantitative nucleoside liquid chrom

SUBMITTER: Dai W 

PROVIDER: S-EPMC8551019 | biostudies-literature | 2021 Nov

REPOSITORIES: biostudies-literature