biostudies-literatureToubdji SDeutsche Forschungsgemeinschaft (DFG)Agence Nationale de la Recherche (ANR)Agence Nationale de la RechercheDeutsche ForschungsgemeinschaftScience Ministry | Forskerakademiet (Danish Research Academy)Animal Health and Welfare ERA-NetAnimal Health and Welfare ERA-Net (ANIHWA)NIGMS NIH HHSScience Ministry | Forskerakademiete2401981121https://www.ebi.ac.uk/biostudies/studies/S-EPMC11317573biostudies-literatureUnknown121(32)Dihydrouridine (D), a prevalent and evolutionarily conserved base in the transcriptome, primarily resides in tRNAs and, to a lesser extent, in mRNAs. Notably, this modification is found at position 2449 in the <i>Escherichia coli</i> 23S rRNA, strategically positioned near the ribosome's peptidyl transferase site. Despite the prior identification, in <i>E. coli</i> genome, of three dihydrouridine synthases (DUS), a set of NADPH and FMN-dependent enzymes known for introducing D in tRNAs and mRNAs, characterization of the enzyme responsible for D2449 deposition has remained elusive. This study introduces a rapid method for detecting D in rRNA, involving reverse transcriptase-blockage at the rhodamine-labeled D2449 site, followed by PCR amplification (RhoRT-PCR). Through analysis of rRNA fromProceedings of the National Academy of Sciences of the United States of AmericaExploring a unique class of flavoenzymes: Identification and biochemical characterization of ribosomal RNA dihydrouridine synthase.PMC11317573GM132254439669440 - TRR-319-TP C01 and HE 3397/21-1FNU-rammebevilling 10-08455420-CE92-0030R01 GM132254Baharoglu ZSudol CHardy LHamdane DGoyenvalle CBregeon DToubdji Sde Crecy-Lagard VThullier QDouthwaite SHelm MMarchand VJean-Jean ORose SKilz LMYuan YMotorin YfalseExploring a unique class of flavoenzymes: Identification and biochemical characterization of ribosomal RNA dihydrouridine synthase.Dihydrouridine (D), a prevalent and evolutionarily conserved base in the transcriptome, primarily resides in tRNAs and, to a lesser extent, in mRNAs. Notably, this modification is found at position 2449 in the <i>Escherichia coli</i> 23S rRNA, strategically positioned near the ribosome's peptidyl transferase site. Despite the prior identification, in <i>E. coli</i> genome, of three dihydrouridine synthases (DUS), a set of NADPH and FMN-dependent enzymes known for introducing D in tRNAs and mRNAs, characterization of the enzyme responsible for D2449 deposition has remained elusive. This study introduces a rapid method for detecting D in rRNA, involving reverse transcriptase-blockage at the rhodamine-labeled D2449 site, followed by PCR amplification (RhoRT-PCR). Through analysis of rRNA from2024-01-01T00:00:00Z2024 Aug2026-05-02T14:33:14.224Z2025-04-04T14:04:35.625ZS-EPMC113175733907867510.1073/pnas.2401981121