Topoisomerase II can unlink replicating DNA by precatenane removal

被引：79

作者：

Lucas, I ^{[1
]}

Germe, T ^{[1
]}

Chevrier-Miller, M ^{[1
]}

Hyrien, O ^{[1
]}

机构：

[1] Ecole Normale Super, Genet Mol Lab, F-75230 Paris 05, France

来源：

EMBO JOURNAL | 2001年 / 20卷 / 22期

关键词：

DNA replication; DNA topoisomerases; nuclear assembly; topoisomerase inhibitors; Xenopus;

D O I：

10.1093/emboj/20.22.6509

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

We have analysed the role of topoisomerase II (topo II) in plasmid DNA replication in Xenopus egg extracts, using specific inhibitors and two-dimensional gel electrophoresis of replication products. Topo II is dispensable for nuclear assembly and complete replication of plasmid DNA but is required for plasmid unlinking. Extensive unlinking can occur in the absence of mitosis. Replication intermediates generated in the absence of topo II activity have an increased positive super-helical stress (+Delta Lk), suggesting a deficiency in pre-catenane removal. The geometry of replication intermediates cut by poisoning topo II with etoposide and purified by virtue of their covalent attachment to topo II subunits demonstrates that topo II acts behind the forks at all stages of elongation. These results provide direct evidence for unlinking replicating DNA by precatenane removal and reveal a division of labour between topo I and topo II in this eukaryotic system. We discuss the role of chromatin structure in driving DNA unlinking during S phase.

引用

页码：6509 / 6519

页数：11

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