Protein architecture of the human kinetochore microtubule attachment site.

Protein architecture of the human kinetochore microtubule attachment site.
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DOI:
10.1016/j.cell.2009.03.035
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发表时间:
2009-05-15
期刊:
影响因子:
64.5
通讯作者:
Salmon ED
Salmon ED
中科院分区:
生物学1区
文献类型:
--
作者:
Wan X;O'Quinn RP;Pierce HL;Joglekar AP;Gall WE;DeLuca JG;Carroll CW;Liu ST;Yen TJ;McEwen BF;Stukenberg PT;Desai A;Salmon ED

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着丝粒介导的着丝粒染色质-纺锤体微管相互作用驱动染色体分离.我们已经开发了一种双色荧光显微镜方法,测量平均标记分离,δ,在< 5 nm的精度-阐明人类中期着丝粒的蛋白质结构。三角洲分析,当与纺锤体连接的姐妹动粒对的张力状态,动粒内的蛋白质连接的机械性能提供了信息。用紫杉醇治疗,它抑制微管动力学,消除紧张的着丝粒,并激活纺锤体检查点,导致特定的大规模的变化,着丝粒结构。累积起来,德尔塔分析显示,符合连接接近着丝粒染色质,建议一个模型,KMN(KNL 1/Mis 12复合物/Ndc 80复合物)网络如何提供微管连接,并从解聚产生拉力,并揭示了紫杉醇治疗诱导的结构变化。这里描述的方法也应该适用于其他中等规模的细胞生物机器。
Centromeric chromatin – spindle microtubule interactions mediated by kinetochores drive chromosome segregation. We have developed a two-color fluorescence light microscopy method that measures average label separation, Delta, at < 5 nm accuracy — to elucidate the protein architecture of human metaphase kinetochores. Delta analysis, when correlated with tension states of spindle-attached sister kinetochore pairs, provided information on mechanical properties of protein linkages within kinetochores. Treatment with taxol—which suppresses microtubule dynamics, eliminates tension at kinetochores, and activates the spindle checkpoint—resulted in specific large-scale changes in kinetochore architecture. Cumulatively, Delta analysis revealed compliant linkages close to the centromeric chromatin, suggests a model for how the KMN (KNL1/Mis12 complex/Ndc80 complex) network provides microtubule attachment and generates pulling forces from depolymerization, and reveals architectural changes induced by taxol treatment. The methods described here should also be applicable to other intermediate-scale biological machines in cells.
DOI: 10.1016/j.cell.2008.03.020
发表时间: 2008-05-02
期刊: CELL
影响因子: 64.5
作者:
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期刊: Current biology : CB
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期刊: CURRENT BIOLOGY
影响因子: 9.2
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