Molecular analysis of core kinetochore composition and assembly in Drosophila melanogaster.
Molecular analysis of core kinetochore composition and assembly in Drosophila melanogaster.
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DOI：
10.1371/journal.pone.0000478
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发表时间：
2007-05-30
期刊：
影响因子：
3.7
通讯作者：
Glover, David M.
中科院分区：
文献类型：
--
作者：
Przewloka, Marcin R.;Zhang, Wei;Costa, Patricia;Archambault, Vincent;D'Avino, Pier Paolo;Lilley, Kathryn S.;Laue, Ernest D.;McAinsh, Andrew D.;Glover, David M.
着丝点是大的多蛋白复合体，是染色体正确分离所必需的。虽然果蝇是染色体分离研究的经典模式生物，但对其着丝点的组织知之甚少。采用生物信息学、蛋白质组学和细胞生物学等方法对果蝇着丝点蛋白相互作用网络进行了鉴定和分析。我们已经证明了三种与人类和酵母高度分化的果蝇蛋白Ndc80、Nuf2和Mis12确实是它们的同源物。从培养的果蝇细胞中对这些蛋白进行亲和纯化，鉴定出另外5种与SPC105/KNL-1、MIND/MIS12和NDC80着丝点复合物亚基弱相似性的相互作用蛋白，以及已知的着丝点相关蛋白，如dynein/dynactin、纺锤体组装检查点组分和异染色质蛋白。在有丝分裂期间，所有8种着丝点复合体蛋白都存在于着丝点上，MIND/MIS12复合体蛋白在间期也存在着丝点。它们的下调导致了染色体聚集/分离的显著缺陷，通常伴随着有丝分裂纺锤体伸长。每个单独的蛋白质的系统耗竭使我们能够建立依赖关系，使它们能够重新聚集到着丝点上。这揭示了首先是MIND/MIS12，然后是NDC80复合体的个体成员的顺序招募。果蝇的MIND/MIS12和NDC80复合体以及Spc105蛋白与其他真核生物的复合体一样，对染色体形成和分离至关重要，但在序列上存在高度分化。个体蛋白的层次依赖关系调节果蝇着丝点复合物的组装，其方式与其他生物体相似，但不完全相同。
Kinetochores are large multiprotein complexes indispensable for proper chromosome segregation. Although Drosophila is a classical model organism for studies of chromosome segregation, little is known about the organization of its kinetochores. We employed bioinformatics, proteomics and cell biology methods to identify and analyze the interaction network of Drosophila kinetochore proteins. We have shown that three Drosophila proteins highly diverged from human and yeast Ndc80, Nuf2 and Mis12 are indeed their orthologues. Affinity purification of these proteins from cultured Drosophila cells identified a further five interacting proteins with weak similarity to subunits of the SPC105/KNL-1, MIND/MIS12 and NDC80 kinetochore complexes together with known kinetochore associated proteins such as dynein/dynactin, spindle assembly checkpoint components and heterochromatin proteins. All eight kinetochore complex proteins were present at the kinetochore during mitosis and MIND/MIS12 complex proteins were also centromeric during interphase. Their down-regulation led to dramatic defects in chromosome congression/segregation frequently accompanied by mitotic spindle elongation. The systematic depletion of each individual protein allowed us to establish dependency relationships for their recruitment onto the kinetochore. This revealed the sequential recruitment of individual members of first, the MIND/MIS12 and then, NDC80 complex. The Drosophila MIND/MIS12 and NDC80 complexes and the Spc105 protein, like their counterparts from other eukaryotic species, are essential for chromosome congression and segregation, but are highly diverged in sequence. Hierarchical dependence relationships of individual proteins regulate the assembly of Drosophila kinetochore complexes in a manner similar, but not identical, to other organisms.
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影响因子：
5.3
作者：
Cortés, A;Azorín, F
通讯作者：
Azorín, F
影响因子：
10.5
作者：
Heeger, S;Leismann, O;Lehner, CF
通讯作者：
Lehner, CF
影响因子：
9.2
作者：
Huertas, D;Cortés, A;Azorín, F
通讯作者：
Azorín, F
DOI：
10.1083/jcb.200210005
发表时间：
2003-01-06
期刊：
The Journal of cell biology
影响因子：
--
作者：
Goshima G;Kiyomitsu T;Yoda K;Yanagida M
通讯作者：
Yanagida M
影响因子：
2.9
作者：
Heit, Ryan;Underhill, D. Alan;Hendzel, Michael J.
通讯作者：
Hendzel, Michael J.