Persistence of DNA threads in human anaphase cells suggests late completion of sister chromatid decatenation.
Persistence of DNA threads in human anaphase cells suggests late completion of sister chromatid decatenation.
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DOI：
10.1007/s00412-007-0131-7
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发表时间：
2008-04
期刊：
影响因子：
1.6
通讯作者：
Nigg EA
中科院分区：
文献类型：
--
作者：
Wang LH;Schwarzbraun T;Speicher MR;Nigg EA
PICH（Plk 1相互作用检查点解旋酶）最近被确定为纺锤体组装检查点的重要组成部分，并显示本地化到着丝粒，内部着丝粒，甚至在分裂后期连接分离染色体的细线。在本文中，我们已经使用免疫荧光原位杂交和染色质免疫沉淀证明PICH与着丝粒染色质在后期。此外，通过仔细分析PICH阳性后期线程通过FISH以及溴脱氧尿苷和CREST标记，我们加强的证据表明，这些线程主要包括α着丝粒脱氧核糖核酸。最后，通过定时添加ICRF-193（拓扑异构酶-II α的特异性抑制剂）的细胞同步在后期，我们证明，拓扑异构酶活性是专门需要解决PICH阳性线程在后期（而不是需要防止形成这样的线程在细胞周期的早期阶段）。这些数据表明，PICH在分裂后期与着丝粒相关，并且大多数PICH阳性丝在内部着丝粒响应张力而伸展时从内部着丝粒进化而来。此外，他们表明，拓扑异构酶的活性是必需的，在分裂后期的决议PICH阳性线程，这意味着完全分离的姐妹染色单体发生晚于以前的假设。本文的在线版本（doi：10.1007/s 00412 -007-0131-7）包含补充材料，可供授权用户使用。
PICH (Plk1-interacting checkpoint helicase) was recently identified as an essential component of the spindle assembly checkpoint and shown to localize to kinetochores, inner centromeres, and thin threads connecting separating chromosomes even during anaphase. In this paper, we have used immuno-fiber fluorescence in situ hybridization and chromatin-immunoprecipitation to demonstrate that PICH associates with centromeric chromatin during anaphase. Furthermore, by careful analysis of PICH-positive anaphase threads through FISH as well as bromo-deoxyurdine and CREST labeling, we strengthen the evidence that these threads comprise mainly alphoid centromere deoxyribonucleic acid. Finally, by timing the addition of ICRF-193 (a specific inhibitor of topoisomerase-II alpha) to cells synchronized in anaphase, we demonstrate that topoisomerase activity is required specifically to resolve PICH-positive threads during anaphase (as opposed to being required to prevent the formation of such threads during earlier cell cycle stages). These data indicate that PICH associates with centromeres during anaphase and that most PICH-positive threads evolve from inner centromeres as these stretch in response to tension. Moreover, they show that topoisomerase activity is required during anaphase for the resolution of PICH-positive threads, implying that the complete separation of sister chromatids occurs later than previously assumed. The online version of this article (doi:10.1007/s00412-007-0131-7) contains supplementary material, which is available to authorized users.
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影响因子：
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