Ndc10 is a platform for inner kinetochore assembly in budding yeast.

Ndc10 is a platform for inner kinetochore assembly in budding yeast.
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DOI:
10.1038/nsmb.2178
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发表时间:
2011-12-04
影响因子:
16.8
通讯作者:
Harrison, Stephen C.
Harrison, Stephen C.
中科院分区:
生物学1区
文献类型:
--
作者:
Cho, Uhn-Soo;Harrison, Stephen C.

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着丝粒将着丝粒DNA连接到纺锤体微管,并确保有丝分裂期间染色体的可靠分离。在点着丝粒酵母中,CBF 3复合物Skp 1:Ctf 13:(Cep 3)2:(Ndc 10)2通过Cep 3和Ndc 10的接触识别保守的着丝粒DNA元件。我们在这里描述的五个结构域组织的乳酸克鲁维酵母Ndc 10和结构域I-II(残基1-402)结合到DNA在2.8 nm分辨率。其结构类似于酪氨酸DNA重组酶,尽管它缺乏内切酶和连接酶活性。结构和生化数据表明,Ndc 10二聚体的每个亚基结合一个单独的DNA片段,这表明Ndc 10稳定在着丝粒的DNA环。我们描述了在体外协会的实验表明,特定的结构域的Ndc 10与每个已知的内动粒蛋白或蛋白复合物在芽殖酵母相互作用。我们认为Ndc 10为内部动粒组装提供了一个中心平台。
Kinetochores link centromeric DNA to spindle microtubules and ensure faithful chromosome segregation during mitosis. In point-centromere yeasts, the CBF3 complex, Skp1:Ctf13:(Cep3)2:(Ndc10)2, recognizes a conserved centromeric DNA element through contacts made by Cep3 and Ndc10. We describe here the five-domain organization of Kluyveromyces lactis Ndc10 and the structure at 2.8 Å resolution of domains I–II (residues 1–402) bound to DNA. The structure resembles tyrosine DNA recombinases, although it lacks both endonuclease and ligase activities. Structural and biochemical data demonstrate that each subunit of the Ndc10 dimer binds a separate fragment of DNA, suggesting that Ndc10 stabilizes a DNA loop at the centromere. We describe in vitro association experiments showing that specific domains of Ndc10 interact with each of the known inner-kinetochore proteins or protein complexes in budding yeast. We propose that Ndc10 provides a central platform for inner-kinetochore assembly.
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