Distinct dynamics of Aurora B and survivin during mitosis
Distinct dynamics of Aurora B and survivin during mitosis
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DOI：
10.4161/cc.3.11.1203
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发表时间：
2004-11-01
期刊：
影响因子：
4.3
通讯作者：
Dimitrov, S
中科院分区：
文献类型：
--
作者：
Delacour-Larose, M;Molla, A;Dimitrov, S
我们研究了极光B和生存素在活细胞有丝分裂过程中的动力学，使用C-末端GFP嵌合体的两种蛋白质。这些嵌合体显示出相同的定位，并表现为真正的野生型蛋白。通过FRAP分析Aurora B-GFP和Survivin-GFP的迁移率。数据显示，与Aurora B-GFP相反，Survivin-GFP在前中期和中期是高度移动的。在分裂末期和细胞分裂时，发现两种嵌合体都是完全不动的。通过siRNA对Aurora B的消融导致Survivin-GFP迁移率的显著降低。这些结果表明，Survivin，而不是极光B，是弱相关的着丝粒染色质在前中期和中期。Survivin与着丝粒染色质的弱关联依赖于Aurora B的存在，并且不受诺考达唑或紫杉醇治疗的影响。乘客蛋白之间的快速和有条件的交换，我们通过实时成像表明，高亲和力的相互作用与乘客蛋白结合的体外分析表明，事实上，静态的“快照”的高度动态和调节的体内相互作用在有丝分裂细胞。
We have studied the dynamics of Aurora B and Survivin during mitosis in living cells, using C-terminal GFP chimeras of the two proteins. These chimeras showed identical localization and behave as bona fide wild type proteins. The mobility of Aurora B-GFP and Survivin-GFP was analyzed by FRAP. The data show that Survivin-GFP, in contrast to Aurora B-GFP, is highly mobile at prometaphase and metaphase. At telophase and cell cleavage, both chimeras are found to be fully immobile. The ablation of Aurora B by siRNA results in a dramatic decrease of the Survivin-GFP mobility. These results demonstrate that Survivin, but not Aurora B, is weakly associated with the centromeric chromatin at prometaphase and metaphase. The weak association of Survivin with centromeric chromatin is dependent on the presence of Aurora B and is not affected by treatment with either nocodazole or taxol. The rapid and conditional interchange between passenger proteins that we show by live imaging indicates that the high affinity interactions demonstrated with in vitro analysis of passenger protein binding are, in fact, static "snapshots" of highly dynamic and regulated in vivo interactions in mitotic cells.