A Dam1-based artificial kinetochore is sufficient to promote chromosome segregation in budding yeast
A Dam1-based artificial kinetochore is sufficient to promote chromosome segregation in budding yeast
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DOI：
10.1038/ncb1924
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发表时间：
2009-09-01
影响因子：
21.3
通讯作者：
Westermann, Stefan
中科院分区：
文献类型：
--
作者：
Kiermaier, Eva;Woehrer, Sophie;Westermann, Stefan
着丝粒是一种大的多蛋白复合物，通过动态连接专门的染色体区域（称为着丝粒）与纺锤体微管的正端来介导所有真核生物中的染色体分离（1，2）。即使是芽殖酵母酿酒酵母的相对简单的动粒也由80多种蛋白质组成，这使得分析它们各自的作用成为一项艰巨的任务（3）。在这里，我们开发了一个系统，使我们能够人工招募蛋白质的DNA序列，并确定它们是否可以提供任何方面的动粒功能在体内。我们表明，人工招聘的微管结合大坝1复合体的质粒缺乏任何着丝粒DNA是足以赋予有丝分裂稳定。基于Dam1的人工动粒能够在有丝分裂纺锤体上附着、双定向和分离微型染色体，并且它们绕过了对天然动粒的必需DNA结合组分的要求。因此，我们已经建立了一个简化的染色体分离系统，直接招募微管力转导组件的DNA。
Kinetochores are large multiprotein complexes that mediate chromosome segregation in all eukaryotes by dynamically connecting specialized chromosome regions, termed centromeres, to the plus-ends of spindle microtubules(1,2). Even the relatively simple kinetochores of the budding yeast Saccharomyces cerevisiae consist of more than 80 proteins, making analysis of their respective roles a daunting task(3). Here, we have developed a system that allows us to artificially recruit proteins to DNA sequences and determine whether they can provide any aspect of kinetochore function in vivo. We show that artificial recruitment of the microtubule-binding Dam 1 complex to a plasmid lacking any centromere DNA is sufficient to confer mitotic stabilization. The Dam1-based artificial kinetochores are able to attach, bi-orient and segregate mini-chromosomes on the mitotic spindle, and they bypass the requirement for essential DNA-binding components of natural kinetochores. Thus, we have built a simplified chromosome segregation system by directly recruiting a microtubule force-transducing component to DNA.