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The first class comprises Nudix hydrolases, including NudC in bacteria,[ 31 , 42 , 43 ] and its eukaryotic homologs, Npy1 in yeast [44] and Nudt12[ 45 , 46 ] and Nudt16 [46] in mammalian cells, which cleave the pyrophosphate bond within the NAD cap to release nicotinamide mononucleotide (NMN) and monophosphorylated RNA (pRNA).[ 31 , 45 , 47 ] The second class includes enzymes from the DXO/Rai1 family, which, unlike Nudix proteins, target the phosphodiester bond to produce pRNAs while releasing an intact NAD molecule.[ 33 , 48 , 49 , 50 ] Notably, homologs of DXO/Rai1 have not been identified in prokaryotes

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