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J Biol Chem, Vol. 273, Issue 50, 33660-33666, December 11, 1998

Essential Mitotic Functions of DNA Topoisomerase IIalpha Are Not Adopted by Topoisomerase IIbeta in Human H69 Cells

Pernille GrueDagger , Alexander GräßerDagger , Maxwell Sehested§, Peter B. Jensen§, Annette UhseDagger , Tobias StraubDagger , Winfried Ness, and Fritz BoegeDagger

From the Dagger  Medizinische Poliklinik, University of Würzburg, Klinikstraße 6-8, D-97070 Würzburg, Germany, the § Department of Pathology and Finsen Center, Rigshospitalet, University of Copenhagen, 2100 Copenhagen, Denmark, and the  Institute of Anatomy, University of Würzburg, D-97070 Würzburg, Germany

Unique functions of mammalian DNA-topoisomerases IIalpha and -beta are suggested by their distinct cellular distribution and chromatin binding at mitosis. Here, we studied H69-VP cells that, due to a homozygous mutation, express topoisomerase IIalpha mostly outside the nucleus. In these cells topoisomerase IIbeta showed a normal nuclear localization. However, at mitosis it diffused away from the chromatin despite the nuclear lack of the alpha -isoform. 80% of these cells performed chromosome condensation and disjunction with the aid of cytosolic topoisomerase IIalpha , which bound to the mitotic chromatin with low affinity. However, the genotype of these cells was highly polyploid indicating an increased rate of non-disjunction. In 20% of the mutant cells neither topoisomerase II isoform was bound to the mitotic chromatin, which appeared as an unstructured DNA spheroid unable to undergo disjunction and cytokinesis. Parental H69 cells expressing topoisomerase IIalpha inside the nucleus exhibited high affinity binding of the enzyme to the mitotic chromatin. Their genotype was mostly diploid and stable. We conclude (i) that high affinity chromatin binding of topoisomerase IIalpha is essential for chromosome condensation/disjunction and (ii) that topoisomerase IIbeta does not adopt these functions.

Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.
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