Dynamic 23S rRNA modification ho5C2501 benefits Escherichia coli under oxidative stress

Fasnacht, Michel; Gallo, Stefano; Sharma, Puneet; Himmelstoß, Maximilian; Limbach, Patrick A; Willi, Jessica; Polacek, Norbert (2022). Dynamic 23S rRNA modification ho5C2501 benefits Escherichia coli under oxidative stress. Nucleic acids research, 50(1), pp. 473-489. Oxford University Press 10.1093/nar/gkab1224

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Post-transcriptional modifications are added to ribosomal RNAs (rRNAs) to govern ribosome biogenesis and to fine-tune protein biosynthesis. In Escherichia coli and related bacteria, RlhA uniquely catalyzes formation of a 5-hydroxycytidine (ho5C) at position 2501 of 23S rRNA. However, the molecular and biological functions as well as the regulation of ho5C2501 modification remain unclear. We measured growth curves with the modification-deficient ΔrlhA strain and quantified the extent of the modification during different conditions by mass spectrometry and reverse transcription. The levels of ho5C2501 in E. coli ribosomes turned out to be highly dynamic and growth phase-dependent, with the most effective hydroxylation yields observed in the stationary phase. We demonstrated a direct effect of ho5C2501 on translation efficiencies in vitro and in vivo. High ho5C2501 levels reduced protein biosynthesis which however turned out to be beneficial for E. coli for adapting to oxidative stress. This functional advantage was small under optimal conditions or during heat or cold shock, but becomes pronounced in the presence of hydrogen peroxide. Taken together, these data provided first functional insights into the role of this unique 23S rRNA modification for ribosome functions and bacterial growth under oxidative stress.

Item Type:

Journal Article (Original Article)


08 Faculty of Science > Department of Chemistry, Biochemistry and Pharmaceutical Sciences (DCBP)

Graduate School:

Graduate School for Cellular and Biomedical Sciences (GCB)

UniBE Contributor:

Fasnacht, Michel, Gallo, Stefano, Sharma, Puneet, Willi, Jessica, Polacek, Norbert


500 Science > 570 Life sciences; biology
500 Science > 540 Chemistry




Oxford University Press




Christina Schüpbach

Date Deposited:

17 Jan 2022 10:08

Last Modified:

05 Dec 2022 15:57

Publisher DOI:


PubMed ID:






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