EB1 and EB3 regulate microtubule minus end organization and Golgi morphology.
EB1 and EB3 regulate microtubule minus end organization and Golgi morphology.
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DOI：
10.1083/jcb.201701024
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发表时间：
2017-10-02
期刊：
影响因子：
--
通讯作者：
Akhmanova A
中科院分区：
文献类型：
--
作者：
Yang C;Wu J;de Heus C;Grigoriev I;Liv N;Yao Y;Smal I;Meijering E;Klumperman J;Qi RZ;Akhmanova A
末端结合蛋白通过募集大量不同的因子来调节微管加末端的动力学和功能。Yang等人表明，EB 1和EB 3也通过参与微管与高尔基体膜的附着而影响微管负末端。该功能对于细胞极性和迁移是重要的。末端结合蛋白（End-binding proteins，EBs）是微管与末端跟踪蛋白复合物的核心组成部分，但目前还不清楚它们是否是哺乳动物微管组织所必需的。在这里，通过使用CRISPR/Cas9介导的敲除技术，我们产生了缺乏EB 2和EB 3以及EB 1的C末端伴侣结合一半的稳定细胞系。这些细胞系在细胞分裂和微管聚合中仅显示出轻度缺陷。然而，在这些细胞中，非中心体微管负端的CAMSAP 2修饰延伸的长度减少，微管与高尔基体膜分离，高尔基体复合体更加紧凑。微管和高尔基体膜的共同组织依赖于EB 1/EB 3-myomegalin复合物，其充当膜微管系链并抵消单个高尔基体堆叠的紧密聚集。EB 1和EB 3的破坏也扰乱细胞迁移、极性和粘着斑的分布。因此，EB 1和EB 3影响多个间期过程，并对微管负末端组织产生重大影响。
End-binding proteins regulate the dynamics and function of microtubule plus ends by recruiting a plethora of diverse factors. Yang et al. show that EB1 and EB3 also affect microtubule minus ends by participating in their attachment to Golgi membranes. This function is important for cell polarity and migration. End-binding proteins (EBs) are the core components of microtubule plus end tracking protein complexes, but it is currently unknown whether they are essential for mammalian microtubule organization. Here, by using CRISPR/Cas9-mediated knockout technology, we generated stable cell lines lacking EB2 and EB3 and the C-terminal partner-binding half of EB1. These cell lines show only mild defects in cell division and microtubule polymerization. However, the length of CAMSAP2-decorated stretches at noncentrosomal microtubule minus ends in these cells is reduced, microtubules are detached from Golgi membranes, and the Golgi complex is more compact. Coorganization of microtubules and Golgi membranes depends on the EB1/EB3–myomegalin complex, which acts as membrane–microtubule tether and counteracts tight clustering of individual Golgi stacks. Disruption of EB1 and EB3 also perturbs cell migration, polarity, and the distribution of focal adhesions. EB1 and EB3 thus affect multiple interphase processes and have a major impact on microtubule minus end organization.
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影响因子：
64.5
作者：
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