Proliferation- and cell cycle-dependent differences in expression of the 170 kilodalton and 180 kilodalton forms of topoisomerase II in NIH-3T3 cells.
The cellular content of 170kD and 180kD topoisomerase II was studied as a function of the proliferation state and cell cycle position in NIH-3T3 cells. When the cells were synchronized by serum starvation and then stimulated to enter the cell cycle by addition of fresh growth medium, the amount of 170kD topoisomerase II present was undetectable until the cells reached late S phase, peaked in G2-M phase cells, and decreased as the cells completed mitosis. The amount of 180kD topoisomerase II was constant once the cells entered the cell cycle. When exponentially growing cells were induced to enter G0 by serum starvation, the amount of 170kD topoisomerase II decreased in parallel with the loss of cells from the S and G2-M phases of the cell cycle and was undetectable once all of the cells reached G0. In contrast, the 180kD enzyme was still present after all of the cells had entered G0. The tightness of association of the two enzymes with chromatin was measured by determining the concentration of salt required to extract them from isolated nuclei. The 180kD enzyme required a higher concentration of NaCl for extraction than did the 170kD enzyme. The different patterns of expression of the two forms of topoisomerase II suggest that they perform different functions in cells.
Woessner
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RD
Research and Development, SmithKline Beecham Pharmaceuticals, King of Prussia, Pennsylvania 19406.
Mattern
M R
MR
Mirabelli
C K
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Johnson
R K
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Drake
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FH
D000818:Animals; D002453:Cell Cycle; D002455:Cell Division; D002478:Cells, Cultured; D002843:Chromatin / Q000378:metabolism; D004250:DNA Topoisomerases, Type II / Q000096:biosynthesis* / Q000235:genetics / Q000502:physiology; D004790:Enzyme Induction; D005347:Fibroblasts / Q000166:cytology / Q000201:enzymology*; D051379:Mice; D008970:Molecular Weight
D002843:Chromatin; D004250:DNA Topoisomerases, Type II