Enzymatic synthesis of polymers containing nicotinamide mononucleotide.
Nicotinamide mononucleoside 5'-diphosphate in its reduced form is an excellent substrate for polynucleotide phosphorylase from Micrococcus luteus both in de novo polymerization reactions and in primer extension reactions. The oxidized form of the diphosphate is a much less efficient substrate; it can be used to extend primers but does not oligomerize in the absence of a primer. The cyanide adduct of the oxidized substrate, like the reduced substrate, polymerizes efficiently. Loss of cyanide yields high molecular weight polymers of the oxidized form. Terminal transferase from calf thymus accepts nicotinamide mononucleoside 5'-triphosphate as a substrate and efficiently adds one residue to the 3'-end of an oligodeoxynucleotide. T4 polynucleotide kinase accepts oligomers of nicotinamide mononucleotide as substrates. However, RNA polymerases do not incorporate nicotinamide mononucleoside 5'-triphosphate into products on any of the templates that we used.
D000818:Animals; D002417:Cattle; D003486:Cyanides / Q000737:chemistry; D018736:DNA Adducts / Q000737:chemistry; D004253:DNA Nucleotidylexotransferase / Q000378:metabolism*; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D011756:Diphosphates / Q000378:metabolism; D016982:Micrococcus luteus / Q000201:enzymology; D009537:Nicotinamide Mononucleotide / Q000378:metabolism*; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis / Q000378:metabolism; D010084:Oxidation-Reduction; D043264:Phosphodiesterase I; D010727:Phosphoric Diester Hydrolases / Q000378:metabolism; D011108:Polymers; D011116:Polynucleotide 5'-Hydroxyl-Kinase / Q000378:metabolism; D011119:Polynucleotides / Q000096:biosynthesis*; D011122:Polyphosphates / Q000378:metabolism; D011117:Polyribonucleotide Nucleotidyltransferase / Q000378:metabolism*; D006163:Ribonuclease T1 / Q000378:metabolism; D012259:Ribonuclease, Pancreatic / Q000378:metabolism; D013379:Substrate Specificity; D013698:Templates, Genetic; D013950:Thymus Gland / Q000201:enzymology
D003486:Cyanides; D018736:DNA Adducts; D011756:Diphosphates; D009838:Oligodeoxyribonucleotides; D011108:Polymers; D011119:Polynucleotides; D011122:Polyphosphates; D009537:Nicotinamide Mononucleotide; D011116:Polynucleotide 5'-Hydroxyl-Kinase; D004253:DNA Nucleotidylexotransferase; D012321:DNA-Directed RNA Polymerases; D011117:Polyribonucleotide Nucleotidyltransferase; D006163:Ribonuclease T1; D012259:Ribonuclease, Pancreatic; D010727:Phosphoric Diester Hydrolases; D043264:Phosphodiesterase I