Identification of NAD-RNA species and ADPR-RNA decapping in Archaea

NAD is a coenzyme central tometabolism that also serves as a 5′-terminal cap for bacterial and eukaryotic transcripts. Thermal degradation of NAD can generate nicotinamide and ADP-ribose (ADPR). Here, we use LC-MS/MS and NAD captureSeq to detect and identify NAD-RNAs in the thermophilic model ar...

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Main Authors: Gomes-Filho, José Vicente, Breuer, Ruth, Morales-Filloy, Hector Gabriel, Pozhydaieva, Nadiia, Borst, Andreas, Paczia, Nicole, Soppa, Jörg, Höfer, Katharina, Jäschke, Andres, Randau, Lennart
Format: Article
Language:English
Published: Philipps-Universität Marburg 2024
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Summary:NAD is a coenzyme central tometabolism that also serves as a 5′-terminal cap for bacterial and eukaryotic transcripts. Thermal degradation of NAD can generate nicotinamide and ADP-ribose (ADPR). Here, we use LC-MS/MS and NAD captureSeq to detect and identify NAD-RNAs in the thermophilic model archaeon Sulfolobus acidocaldarius and in the halophilic mesophile Haloferax volcanii. None of the four Nudix proteins of S. acidocaldarius catalyze NADRNA decapping in vitro, but one of the proteins (Saci_NudT5) promotes ADPRRNA decapping. NAD-RNAs are converted into ADPR-RNAs, which we detect in S. acidocaldarius total RNA. Deletion of the gene encoding the 5′−3′ exonuclease Saci-aCPSF2 leads to a 4.5-fold increase in NAD-RNA levels.We propose that the incorporation of NAD into RNA acts as a degradation marker for SaciaCPSF2. In contrast, ADPR-RNA is processed by Saci_NudT5 into 5′-p-RNAs, providing another layer of regulation for RNA turnover in archaeal cells.
Item Description:Gefördert durch den Open-Access-Publikationsfonds der UB Marburg.
DOI:10.1038/s41467-023-43377-x