Nucleic Acids Research, 1995, Vol. 23, No. 18 3742-3749
© 1995
ENZYMOLOGY |
Enzymatic synthesis of polymers containing nicotinamide mononucleotide
The Salk Institute for Biological Studies PO Box 85800, San Diego, CA 92186-5800, USA
*To whom correspondence should be addressed
Received May 15, 1995. Revised August 9, 1995. Accepted August 9, 1995.
Nicotinamide mononucleoside 5'-diphosphate in its reduced form is an excellent substrate for polynucieotide 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'-triphos-phate into products on any of the templates that we used.