RNAi系统分析染色体分离蛋白

【摘要】 奥地利分子病理学研究所，德国Max Planck研究所分子细胞生物与遗传学实验室，Wellcome Trust Sanger研究所等多国的科研工作者在人类染色体分离蛋白研究方面取得新进展，相关成果文章Systematic Analysis of Human Protein Complexes Identifies Chromosome Segregation Proteins公布在Science在线版上。

染色体分离与细胞分裂是关系生命体生存的关键生命活动过程，在这2个过程中有大量的蛋白复合物参与调控。

近期，多国研究人员组成的联合研究小组应用RNAi技术对大量参与其中的蛋白质复合物进行扫描分析，整个过程鉴定了许多与染色体分离、细胞分裂相关的蛋白复合物，但是，关于这些蛋白复合物的功能研究却十分罕见。

为了探明这些蛋白复合物，科学家们应用基因标签技术，蛋白定位技术、MitoCheck等技术分析100种人类蛋白复合物，这100种蛋白复合物大部分的还没有被全面的了解。这项新的研究工作将在前人的基础上，对这100个蛋白复合物进行全面的分析。

这种高通量的分析有助人们了解蛋白复合物的亚基、复合体等多个结构的生理功能。

关于染色体分离

染色体分离是有丝分裂和减数分裂的关键事件，是保证姊妹(同源)染色体正确分配到子细胞中关键的调控环节之一。近年来，利用酵母等真核模式生物突变体，研究人员克隆了一些调控细胞分裂早期姊妹染色体黏着和后期黏着素酶解的关键基因,初步阐明了染色体分离的分子调控机理。由多亚基蛋白组成的黏着素是保证染色体正确黏着和分离的关键分子。在细胞分裂中后期过渡时，经过一系列的分子识别和互作导致黏着素蛋白选择性地降解，使染色体分离。

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Systematic Analysis of Human Protein Complexes Identifies Chromosome Segregation Proteins

James R.A. Hutchins,1~undefined Yusuke Toyoda,2~undefined Björn Hegemann,1~undefined, Ina Poser,2~undefined Jean-Karim Hériché,3,4 Martina M. Sykora,1 Martina Augsburg,2 Otto Hudecz,1 Bettina A. Buschhorn,1 Jutta Bulkescher,4 Christian Conrad,4 David Comartin,5,6 Alexander Schleiffer,1 Mihail Sarov,2 Andrei Pozniakovsky,2 Mikolaj Michal Slabicki,2 Siegfried Schloissnig,2,7 Ines Steinmacher,1 Marit Leuschner,2 Andrea Ssykor,2 Steffen Lawo,5,6 Laurence Pelletier,5,6 Holger Stark,8 Kim Nasmyth,1, Jan Ellenberg,4 Richard Durbin,3 Frank Buchholz,2 Karl Mechtler,1 Anthony A. Hyman,2, Jan-Michael Peters1,

Chromosome segregation and cell division are essential, highly ordered processes that depend on numerous protein complexes. Results from recent RNA interference (RNAi) screens indicate that the identity and composition of these protein complexes is incompletely understood. Using gene tagging on bacterial artificial chromosomes, protein localization and tandem affinity purification-mass spectrometry, the MitoCheck consortium has analyzed about 100 human protein complexes, many of which had not or only incompletely been characterized. This work has led to the discovery of previously unknown, evolutionarily conserved subunits of the anaphase-promoting complex (APC/C) and the -tubulin ring complex (-TuRC), large complexes which are essential for spindle assembly and chromosome segregation. The approaches we describe here are generally applicable to high throughput follow-up analyses of phenotypic screens in mammalian cells.

1Research Institute of Molecular Pathology, Dr. Bohr-Gasse 7, A-1030 Vienna, Austria.

2Max Planck Institute for Molecular Cell Biology and Genetics, Pfotenhauerstrasse 108, D-01307 Dresden, Germany.

3Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge, CB10 1HH, UK.

4Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Meyerhofstrasse 1, D-69117 Heidelberg, Germany.

5Samuel Lunenfeld Research Institute, Mount Sinai Hospital, 600 University Avenue, Toronto, Ontario, M5G 1X5, Canada.

6Department of Molecular Genetics, University of Toronto, Toronto, Ontario, M5S 1A8, Canada.

7German Cancer Research Center, Im Neuenheimer Feld 280, 69120 Heidelberg, Germany.

8Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, D-37077 Göttingen, Germany.