Dynamic behavior of DNA topoisomerase IIβ in response to DNA double-strand breaks.
Dynamic behavior of DNA topoisomerase IIβ in response to DNA double-strand breaks.
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DOI：
10.1038/s41598-018-28690-6
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发表时间：
2018-07-09
影响因子：
4.6
通讯作者：
Yano KI
中科院分区：
文献类型：
--
作者：
Morotomi-Yano K;Saito S;Adachi N;Yano KI
DNA拓扑异构酶II(Topo II)对于解决DNA的拓扑结构问题至关重要，在复制、转录和染色体分离等多种细胞过程中发挥着重要作用。虽然在DNA修复过程中也可能出现DNA拓扑问题，但Topo II在这一过程中的可能参与仍有待充分调查。在这里，我们展示了人类TOPO IIβ对DNA双链断裂(DSB)的动态响应行为，双链断裂是DNA损伤的最有害形式。活细胞成像结合激光微照射定点诱导DSB，显示了绿色荧光蛋白标记的TOPO IIβ快速募集到DSB位点。洗涤剂提取和免疫荧光显示内源性TOPO IIβ与DSB位点紧密结合。光漂白分析表明TOPO IIβ在细胞核内具有很高的流动性。TOPO II催化抑制剂ICRF-187和ICRF-193降低了TOPO II的β迁移率，从而阻止了TOPO IIβ向DSB的募集。此外，TOPO IIβ基因敲除细胞对博莱霉素的敏感性增加，同源重组(HR)介导的DSB修复减少，提示TOPO IIβ在HR介导的DSB修复中起作用。综上所述，这些结果突出了TOPO IIβ功能在细胞对DSB反应中的一个新方面。
DNA topoisomerase II (Topo II) is crucial for resolving topological problems of DNA and plays important roles in various cellular processes, such as replication, transcription, and chromosome segregation. Although DNA topology problems may also occur during DNA repair, the possible involvement of Topo II in this process remains to be fully investigated. Here, we show the dynamic behavior of human Topo IIβ in response to DNA double-strand breaks (DSBs), which is the most harmful form of DNA damage. Live cell imaging coupled with site-directed DSB induction by laser microirradiation demonstrated rapid recruitment of EGFP-tagged Topo IIβ to the DSB site. Detergent extraction followed by immunofluorescence showed the tight association of endogenous Topo IIβ with DSB sites. Photobleaching analysis revealed that Topo IIβ is highly mobile in the nucleus. The Topo II catalytic inhibitors ICRF-187 and ICRF-193 reduced the Topo IIβ mobility and thereby prevented Topo IIβ recruitment to DSBs. Furthermore, Topo IIβ knockout cells exhibited increased sensitivity to bleomycin and decreased DSB repair mediated by homologous recombination (HR), implicating the role of Topo IIβ in HR-mediated DSB repair. Taken together, these results highlight a novel aspect of Topo IIβ functions in the cellular response to DSBs.
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DOI：
10.1083/jcb.200510130
发表时间：
2006-04-24
期刊：
The Journal of cell biology
影响因子：
--
作者：
Bekker-Jensen S;Lukas C;Kitagawa R;Melander F;Kastan MB;Bartek J;Lukas J
通讯作者：
Lukas J
影响因子：
7.8
作者：
Kim, Jong-Soo;Krasieva, Tatiana B;Kurumizaka, Hitoshi;Chen, David J;Taylor, A Malcolm R;Yokomori, Kyoko
通讯作者：
Yokomori, Kyoko
影响因子：
64.8
作者：
Berger, JM;Gamblin, SJ;Wang, JC
通讯作者：
Wang, JC
DOI：
10.1016/0167-4781(92)90050-a
发表时间：
1992-08-17
期刊：
BIOCHIMICA ET BIOPHYSICA ACTA
影响因子：
--
作者：
CAPRANICO, G;TINELLI, S;ZUNINO, F
通讯作者：
ZUNINO, F
XRCC1募集对单链断裂和基础切除修复的不同时空模式和PARP依赖性。
影响因子：
14.9
作者：
Campalans A;Kortulewski T;Amouroux R;Menoni H;Vermeulen W;Radicella JP
通讯作者：
Radicella JP