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First published online March 2, 2004
doi: 10.1242/10.1242/jcs.00977

Journal of Cell Science 117, 1047-1054 (2004)
Published by The Company of Biologists 2004
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Research Article

Functional compatibility between isoform {alpha} and ß of type II DNA topoisomerase

Ayako Sakaguchi* and Akihiko Kikuchi{ddagger}

Laboratory of Medical Mycology, Research Institute for Disease Mechanism and Control, Nagoya University Graduate School and Faculty of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya 466-8550, Japan

{ddagger} Author for correspondence (e-mail: aki{at}med.nagoya-u.ac.jp)

Accepted 11 November 2003

DNA topoisomerase II (topo II) plays a crucial role in controlling the conformation of both DNA and whole chromosomes. This activity is essential for several cellular events such as DNA replication, transcription, chromosome condensation and segregation. In mammals, two genes code for isoforms of topo II, termed {alpha} and ß. They are similar in primary structure and have almost identical catalytic properties in vitro. We transfected HeLa cells with small interfering RNAs (siRNAs) targeted against either topo II{alpha} or IIß, and succeeded in knocking down the expression of the corresponding protein. Chromosomes were condensed and aligned at metaphase in topo II{alpha}-knockdown cells. Although some lagging chromosomes were observed, they were still segregated at anaphase despite the absence of topo II{alpha}. When both topo II{alpha} and topo II{gamma} were removed, the segregation of chromosomes was severely arrested, suggesting that topo II{gamma} could partially substitute for topo II{alpha}. Double-knockdown experiments also revealed that topo II was required for shortening of the chromosome axis.

Key words: Chromosome condensation, Chromosome segregation, DNA topoisomerase II{alpha}, DNA topoisomerase IIß, siRNA




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