A novel NAD-RNA decapping pathway discovered by synthetic light-up NAD-RNAs

• Verfasst von: Abele, Florian [VerfasserIn]
• Verfasst von: Höfer, Katharina [VerfasserIn]
• Verfasst von: Bernhard, Patrick [VerfasserIn]
• Verfasst von: Grawenhoff, Julia [VerfasserIn]
• Verfasst von: Seidel, Maximilian [VerfasserIn]
• Verfasst von: Krause, André [VerfasserIn]
• Verfasst von: Kopf, Sara [VerfasserIn]
• Verfasst von: Schröter, Martin [VerfasserIn]
• Verfasst von: Jäschke, Andres [VerfasserIn]
• Titel: A novel NAD-RNA decapping pathway discovered by synthetic light-up NAD-RNAs
• Verf.angabe: Florian Abele, Katharina Höfer, Patrick Bernhard, Julia Grawenhoff, Maximilian Seidel, André Krause, Sara Kopf, Martin Schröter and Andres Jäschke
• E-Jahr: 2020
• Jahr: 28 March 2020
• Umfang: 15 S.
• Fussnoten: Gesehen am 02.07.2020
• Titel Quelle: Enthalten in: Biomolecules
• Ort Quelle: Basel : MDPI, 2011
• Jahr Quelle: 2020
• Band/Heft Quelle: 10(2020), 4, Artikel-ID 513, Seite 1-15
• ISSN Quelle: 2218-273X
• DOI: doi:10.3390/biom10040513
• Datenträger: Online-Ressource
• Sprache: eng
• Sach-SW: Decapping
• Sach-SW: fluorescent RNA
• Sach-SW: glycohydrolase
• Sach-SW: high-throughput screening
• Sach-SW: NAD metabolism
• Sach-SW: NAD-capped RNA
• Sach-SW: RNA modification
• K10plus-PPN: 1703276027

Abstract

The complexity of the transcriptome is governed by the intricate interplay of transcription, RNA processing, translocation, and decay. In eukaryotes, the removal of the 5’-RNA cap is essential for the initiation of RNA degradation. In addition to the canonical 5’-N7-methyl guanosine cap in eukaryotes, the ubiquitous redox cofactor nicotinamide adenine dinucleotide (NAD) was identified as a new 5’-RNA cap structure in prokaryotic and eukaryotic organisms. So far, two classes of NAD-RNA decapping enzymes have been identified, namely Nudix enzymes that liberate nicotinamide mononucleotide (NMN) and DXO-enzymes that remove the entire NAD cap. Herein, we introduce 8-(furan-2-yl)-substituted NAD-capped-RNA (FurNAD-RNA) as a new research tool for the identification and characterization of novel NAD-RNA decapping enzymes. These compounds are found to be suitable for various enzymatic reactions that result in the release of a fluorescence quencher, either nicotinamide (NAM) or nicotinamide mononucleotide (NMN), from the RNA which causes a fluorescence turn-on. FurNAD-RNAs allow for real-time quantification of decapping activity, parallelization, high-throughput screening and identification of novel decapping enzymes in vitro. Using FurNAD-RNAs, we discovered that the eukaryotic glycohydrolase CD38 processes NAD-capped RNA in vitro into ADP-ribose-modified-RNA and nicotinamide and therefore might act as a decapping enzyme in vivo. The existence of multiple pathways suggests that the decapping of NAD-RNA is an important and regulated process in eukaryotes.