Growth state and cell cycle dependent phosphorylation of DNA topoisomerase II in Swiss 3T3 cells.
We have investigated the amount of DNA topoisomerase II and phosphorylation of the enzyme in Swiss 3T3 cells during the transition from cell quiescence to proliferation. A relatively high level of phosphorylation was observed with proliferating cells while no or a very low level of phosphorylation was observed with quiescent cells. Phosphoamino acid analysis of the phosphorylated topoisomerase II revealed that the phosphorylated aminoacyl residue was serine. When quiescent cells were stimulated to grow by the addition of serum, DNA synthesis began to increase at 9 h after serum addition, reaching a maximum at 15 h and then declining. The amount of topoisomerase II began to increase at 6 h and reached a maximum at 22-27 h, corresponding to the G2 phase. The phosphorylation of topoisomerase II measured by pulse-labeling gradually increased from 6 to 18 h and reached a maximum at 22 h when the amount of the enzyme was maximum. The level of phosphorylation measured by continuous-labeling increased gradually up to 12 h and markedly up to 28 h, and then declined. The increase in the rate of phosphorylation in the G2 phase was affected by inhibiting DNA synthesis, but the increase in the amount of the enzyme was not. Thus, it was suggested that the regulation of phosphorylation of topoisomerase II differs from that of the amount of the enzyme.
Saijo
M
M
Department of Physiological Chemistry, Faculty of Pharmaceutical Sciences, University of Tokyo, Japan.
Ui
M
M
Enomoto
T
T
D016475:3T3 Cells; D000818:Animals; D016590:Aphidicolin / Q000494:pharmacology; D002453:Cell Cycle; D002455:Cell Division; D004247:DNA / Q000037:antagonists & inhibitors / Q000096:biosynthesis; D004250:DNA Topoisomerases, Type II / Q000276:immunology / Q000378:metabolism*; D007106:Immune Sera / Q000502:physiology; D007399:Interphase; D051379:Mice; D010766:Phosphorylation; D011817:Rabbits
D007106:Immune Sera; D016590:Aphidicolin; D004247:DNA; D004250:DNA Topoisomerases, Type II