ABSTRACT
The biochemical activity of terminal transferase (TdT) in the thymocytes of leukemic AKR mice has no relationship to cell cycle stage, unlike the activity of replicative DNA polymerase which increases during the period of DNA synthesis. Moreover, such assays of DNA polymerase alpha reveal a shift in enzyme activity from cytoplasm to nucleus during S phase. In the present study, the role of TdT in DNA metabolism was explored further by examining the intracellular location of the enzyme during cytokinesis. Single cell suspensions of thymocytes from leukemic AKR mice were partially synchronized by velocity sedimentation in a sucrose gradient at unit gravity and harvested according to cell cycle stage. The content and location of TdT in individual cells was determined by indirect immunofluorescence using a rabbit antiserum to calf thymus TdT as the primary antibody. There was no relationship of fluorescence intensity or of the proportion of TdT-positive cells to cell cycle stage. In all samples examined (n = 6) the enzyme was located almost entirely in the nucleus throughout cytokinesis. These results do not support the hypothesis that the intracellular location of TdT may vary with cell cycle stage and a role for the enzyme in DNA synthesis remains to be defined.
