The yeast DASH complex forms closed rings on microtubules
The yeast DASH complex forms closed rings on microtubules
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DOI：
10.1038/nsmb896
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发表时间：
2005-02-01
影响因子：
16.8
通讯作者：
Harrison, SC
中科院分区：
文献类型：
--
作者：
Miranda, JJL;De Wulf, P;Harrison, SC
酿酒酵母DASH复合体是动粒微管结合的重要组成部分。我们在大肠杆菌中共表达了该组件的所有10个亚基，并纯化了一个单一的复合体，即210 kDa的异构体，每个亚基的一个拷贝具有明显的化学计量比。重组组件的流体力学性质与酵母抽提物中的天然复合体没有区别。光镜下观察DASH与微管结合的结构。游离的杂十聚体相对球状。在微管存在的情况下，DASH寡聚形成环和围绕微管的成对螺旋。我们讨论了这种领状结构在染色体分离过程中维持微管附着和纺锤体完整性方面的潜在作用。
The Saccharomyces cerevisiae DASH complex is an essential microtubule- binding component of the kinetochore. We coexpressed all ten subunits of this assembly in Escherichia coli and purified a single complex, a 210-kDa heterodecamer with an apparent stoichiometry of one copy of each subunit. The hydrodynamic properties of the recombinant assembly are indistinguishable from those of the native complex in yeast extracts. The structure of DASH alone and bound to microtubules was visualized by EM. The free heterodecamer is relatively globular. In the presence of microtubules, DASH oligomerizes to form rings and paired helices that encircle the microtubules. We discuss potential roles for such collar-like structures in maintaining microtubule attachment and spindle integrity during chromosome segregation.