Sakaguchi Ayako, Kikuchi Akihiko
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.
J Cell Sci. 2004 Mar 1;117(Pt 7):1047-54. doi: 10.1242/jcs.00977.
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 beta. 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 IIalpha or IIbeta, and succeeded in knocking down the expression of the corresponding protein. Chromosomes were condensed and aligned at metaphase in topo IIalpha-knockdown cells. Although some lagging chromosomes were observed, they were still segregated at anaphase despite the absence of topo IIalpha. When both topo IIalpha and topo IIbeta were removed, the segregation of chromosomes was severely arrested, suggesting that topo IIbeta could partially substitute for topo IIalpha. Double-knockdown experiments also revealed that topo II was required for shortening of the chromosome axis.
DNA拓扑异构酶II(拓扑异构酶II)在控制DNA和整个染色体的构象方面起着至关重要的作用。这种活性对于诸如DNA复制、转录、染色体浓缩和分离等多种细胞事件至关重要。在哺乳动物中,有两个基因编码拓扑异构酶II的同工型,称为α和β。它们的一级结构相似,在体外具有几乎相同的催化特性。我们用针对拓扑异构酶IIα或IIβ的小干扰RNA(siRNA)转染了HeLa细胞,并成功敲低了相应蛋白质的表达。在拓扑异构酶IIα敲低的细胞中,染色体在中期浓缩并排列。尽管观察到一些滞后染色体,但尽管没有拓扑异构酶IIα,它们在后期仍能分离。当拓扑异构酶IIα和IIβ都被去除时,染色体的分离被严重阻滞,这表明拓扑异构酶IIβ可以部分替代拓扑异构酶IIα。双敲低实验还表明,拓扑异构酶II是染色体轴缩短所必需的。