J. Cell Biol.
© The Rockefeller University Press
0021-9525/97/02/775/14 $2.00
Volume 136, Number 4, February 24, 1997 775-788

Cell Cycle-coupled Relocation of Types I and II Topoisomerases and Modulation of Catalytic Enzyme Activities

Kay N. Meyer,^* Eigil Kjeldsen,^§ Tobias Straub,^* Birgitta R. Knudsen, Ian D. Hickson,^¶ Akihiko Kikuchi,^** Hans Kreipe, and Fritz Boege^*

^* Medizinische Poliklinik and  Institute of Pathology, University of Würzburg, Germany; ^§ Department of Human Genetics and  Department of Molecular and Structural Biology, University of Aarhus, Denmark; ^¶ Institute of Molecular Medicine, John Radcliffe Hospital, University of Oxford, United Kingdom; and ^** Laboratory of Medical Mycology, Institute for Disease Mechanism and Control, Nagoya University, Japan

We visualized DNA topoisomerases in A431 cells and isolated chromosomes by isoenzyme-selective immunofluorescence microscopy. In interphase, topoisomerase I mainly had a homogeneous nuclear distribution. 10-15% of the cells exhibited granular patterns, 30% showed bright intranucleolar patches. Topoisomerase II isoenzymes showed spotted () or reticular () nuclear patterns throughout interphase. In contrast to topoisomerase II, topoisomerase II was completely excluded from nucleoli. In mitosis, topoisomerase II diffused completely into the cytosol, whereas topoisomerases I and II remained chromosome bound. Chromosomal staining of topoisomerase I was homogeneous, whereas topoisomerase II accumulated in the long axes of the chromosome arms and in the centriols. Topoisomerase antigens were 2-3-fold higher in mitosis than in interphase, but specific activities of topoisomerase I and II were reduced 5- and 2.4-fold, respectively. These changes were associated with mitotic enzyme hyperphosphorylation. In interphase, topoisomerases could be completely linked to DNA by etoposide or camptothecin, whereas in mitosis, 50% of topoisomerase II escaped poisoning. Refractoriness to etoposide could be assigned to the salt-stable scaffold fraction of topoisomerase II, which increased from <2% in G₁ phase to 48% in mitosis. Topoisomerases I and II remained completely extractable throughout the cell cycle. In summary, expression of topoisomerases increases towards mitosis, but specific activities decrease. Topoisomerase II is released from the heterochromatin, whereas topoisomerase I and II remain chromosome bound. Scaffold-associated topoisomerase II appears not to be involved in catalytic DNA turnover, though it may play a role in the replicational cycle of centriols, where it accumulates during M phase.

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