Role of survivin phosphorylation by Aurora B in mitosis
Role of survivin phosphorylation by Aurora B in mitosis
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DOI：
10.4161/cc.6.15.4482
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发表时间：
2007-08-01
期刊：
影响因子：
4.3
通讯作者：
Molla, Annie
中科院分区：
文献类型：
--
作者：
Delacour-Larose, Marlene;Thi, My-Nhung Hoang;Molla, Annie
染色体蛋白乘客复合物是一种关键的有丝分裂调节因子，由至少四种蛋白质组成，INCENP、Aurora B、Survivin和Borealin。与染色体蛋白乘客复合物（CPC）的其他成员相反，存活素在中期是移动的。该蛋白质也被Aurora B在Three 117处磷酸化。在这项工作中，我们已经研究了磷酸化的Survivin在有丝分裂中的作用，通过使用非磷酸化的T117 A和磷酸化的T117 E沉默抗性Survivin突变体，表达这些突变体的诱导细胞系和siRNA，延时显微镜和FRAP分析的组合。时间推移显微镜和FRAP分析表明，生存素T117 A突变体是非常稳定的着丝粒和它的表达诱导内源性生存素耗尽细胞的前中期阻滞。此外，生存素T117 A无法拯救内源性生存素缺失细胞的表型。在这些细胞中表达，磷酸化模拟生存素T117 E突变体表现出非常弱的相互作用与着丝粒和行为作为一个显性负突变体诱导严重的有丝分裂缺陷。我们的数据表明，极光B产生的磷酸化/去磷酸化周期的生存是必需的正常进行的有丝分裂。
The chromosomal protein passenger complex, a key mitotic regulator, consists of at least four proteins, INCENP, Aurora B, Survivin and Borealin. Survivin, in contrast to the other members of the chromosomal protein passenger complex (CPC), is mobile at metaphase. This protein is also phosphorylated by Aurora B at Threonine 117. In this work we have studied the role of the phosphorylation of Survivin in mitosis by using non phosphorylable T117A and phosphomimic T117E silent resistant Survivin mutants, inducible cell lines expressing these mutants and a combination of siRNA, time-lapse microscopy and FRAP analysis. Time lapse microscopy and FRAP analysis show that Survivin T117A mutant is very stably associated with centromeres and its expression induces a prometaphasic arrest in endogenous survivin depleted cells. In addition, Survivin T117A was unable to rescue the phenotypes of the endogenous survivin depleted cells. Expressed in these cells, the phosphomimic Survivin T117E mutant exhibits a very weak interaction with the centromeres and behaves as a dominant negative mutant inducing severe mitotic defects. Our data suggest that the Aurora B generated phosphorylation/dephosphorylation cycle of Survivin is required for proper proceeding of mitosis.