Augmin-dependent microtubule nucleation at microtubule walls in the spindle.

Augmin-dependent microtubule nucleation at microtubule walls in the spindle.
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DOI:
10.1083/jcb.201304031
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发表时间:
2013-07-08
期刊:
The Journal of cell biology
影响因子:
--
通讯作者:
Goshima G
Goshima G
中科院分区:
其他
文献类型:
--
作者:
Kamasaki T;O'Toole E;Kita S;Osumi M;Usukura J;McIntosh JR;Goshima G

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电子断层扫描和 3D 建模可识别一根微管壁与纺锤体中相邻微管负端之间依赖于奥格蛋白的连接。功能纺锤体的形成需要纺锤体内部的微管(MT)成核,这取决于奥格明。 augmin 如何促进 MT 形成和组织尚不清楚,因为依赖 augmin 的 MT 从未被具体可视化。在本文中,我们通过电子断层扫描和 3D 建模识别了依赖于 augmin 的 MT 及其与其他 MT 的连接。在人类细胞的中期纺锤体中,MT 的负端既位于中心粒周围,又位于纺锤体内部。当augmin被敲低时,后者的MT数量显着减少。在对照细胞中,我们确定了一个 MT 的壁与相邻 MT 负端之间的连接。有趣的是,与细胞骨架聚合物之间端壁连接的其他例子不同,连接的 MT 几乎是平行的。我们的观察结果支持了现有纺锤体 MT 壁上依赖于 Augmin 的 MT 成核的概念。此外,他们提出了一种维持极化 MT 组织的机制,即使非中心体 MT 起始广泛存在。
Electron tomography and 3D modeling identifies augmin-dependent connections between the wall of one microtubule and the minus end of a neighboring one in the spindle. The formation of a functional spindle requires microtubule (MT) nucleation from within the spindle, which depends on augmin. How augmin contributes to MT formation and organization is not known because augmin-dependent MTs have never been specifically visualized. In this paper, we identify augmin-dependent MTs and their connections to other MTs by electron tomography and 3D modeling. In metaphase spindles of human cells, the minus ends of MTs were located both around the centriole and in the body of the spindle. When augmin was knocked down, the latter population of MTs was significantly reduced. In control cells, we identified connections between the wall of one MT and the minus end of a neighboring MT. Interestingly, the connected MTs were nearly parallel, unlike other examples of end–wall connections between cytoskeletal polymers. Our observations support the concept of augmin-dependent MT nucleation at the walls of existing spindle MTs. Furthermore, they suggest a mechanism for maintaining polarized MT organization, even when noncentrosomal MT initiation is widespread.
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