Properties of a terminal deoxyribonucleotidyltransferase isolated from wheat germ.

An enzyme able to catalyse the polymerization of deoxyribonucleotides in a template-independent manner was isolated from dry wheat germ. This activity is associated with a soluble protein which is homogeneous with respect to the molecular weight (approx. 500000) and, under denaturing conditions, dissociates into product of two size classes, 67000 and 45000 daltons respectively. The enzyme-catalysed polymerization can be primed by oligo- as well as poly-deoxyribonucleotides, and is highly efficient (234 nmol/h per mg of finally purified protein) when only one of the four deoxyribonucleoside 5'-triphosphates is present in the incubation mixture. An extension of the 3'-hydroxy termini of polydeoxyribonucleotide chains for approx. 40 nucleotide residues was achieved when non-denatured DNA and [3H]dTTP were used as the primer and substrate respectively. It is concluded that the enzyme isolated from wheat germ shares catalytic properties with the terminal deoxynucleotidyltransferase of mammalian thymus. Unlike that transferase, however, the plant enzyme prefers non-denatured to single-stranded DNA as primer and requires both Mg2+ and Mn2+ ions for maximal activity.