AI Article Synopsis

  • Scientists found that RNA can be changed in special ways that affect how it works, but they don't know a lot about this yet.
  • They created a new method called RNABPP to study how these changes happen in human cells.
  • Their research shows that when a certain protein (DUS3L) is missing, cells can't make proteins properly and don't grow as they should, which helps us learn more about how RNA modifications work.

Article 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 (mC) and 5-methyluridine (mU) 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 chromatography-tandem mass spectrometry (LC-MS/MS) analysis and 5-FUrd-based crosslinking and immunoprecipitation (CLIP) sequencing to map DUS3L-dependent dihydrouridine (DHU) modifications across the transcriptome. Finally, we show that DUS3L-knockout (KO) cells have compromised protein translation rates and impaired cellular proliferation. Taken together, our work provides a general approach for profiling RNA-modifying enzyme activity in living cells and reveals new pathways for epitranscriptomic RNA regulation.

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8551019PMC
http://dx.doi.org/10.1038/s41589-021-00874-8DOI Listing

Publication Analysis

Top Keywords

rna-modifying enzyme
8
epitranscriptomic rna
8
rna
6
activity-based rna-modifying
4
enzyme probing
4
probing reveals
4
reveals dus3l-mediated
4
dus3l-mediated dihydrouridylation
4
dihydrouridylation epitranscriptomic
4
rna modifications
4

Similar Publications

Want AI Summaries of new PubMed Abstracts delivered to your In-box?

Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!