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细胞周期蛋白依赖性激酶11(Cdk11)是一种依赖于RanGTP的微管稳定因子,可调节纺锤体组装速率。

Cdk11 is a RanGTP-dependent microtubule stabilization factor that regulates spindle assembly rate.

作者信息

Yokoyama Hideki, Gruss Oliver J, Rybina Sofia, Caudron Maïwen, Schelder Malgorzata, Wilm Matthias, Mattaj Iain W, Karsenti Eric

机构信息

European Molecular Biology Laboratory, Heidelberg 69117, Germany.

出版信息

J Cell Biol. 2008 Mar 10;180(5):867-75. doi: 10.1083/jcb.200706189. Epub 2008 Mar 3.

Abstract

Production of Ran-guanosine triphosphate (GTP) around chromosomes induces local nucleation and plus end stabilization of microtubules (MTs). The nuclear protein TPX2 is required for RanGTP-dependent MT nucleation. To find the MT stabilizer, we affinity purify nuclear localization signal (NLS)-containing proteins from Xenopus laevis egg extracts. This NLS protein fraction contains the MT stabilization activity. After further purification, we used mass spectrometry to identify proteins in active fractions, including cyclin-dependent kinase 11 (Cdk11). Cdk11 localizes on spindle poles and MTs in Xenopus culture cells and egg extracts. Recombinant Cdk11 demonstrates RanGTP-dependent MT stabilization activity, whereas a kinase-dead mutant does not. Inactivation of Cdk11 in egg extracts blocks RanGTP-dependent MT stabilization and dramatically decreases the spindle assembly rate. Simultaneous depletion of TPX2 completely inhibits centrosome-dependent spindle assembly. Our results indicate that Cdk11 is responsible for RanGTP-dependent MT stabilization around chromosomes and that this local stabilization is essential for normal rates of spindle assembly and spindle function.

摘要

染色体周围的Ran-鸟苷三磷酸(GTP)生成诱导微管(MTs)的局部成核和正端稳定。核蛋白TPX2是RanGTP依赖性MT成核所必需的。为了找到MT稳定剂,我们从非洲爪蟾卵提取物中亲和纯化含核定位信号(NLS)的蛋白质。这个NLS蛋白组分含有MT稳定活性。进一步纯化后,我们用质谱法鉴定活性组分中的蛋白质,包括细胞周期蛋白依赖性激酶11(Cdk11)。Cdk11定位于非洲爪蟾培养细胞和卵提取物中的纺锤体极和MTs上。重组Cdk11表现出RanGTP依赖性MT稳定活性,而激酶失活突变体则没有。卵提取物中Cdk11的失活阻断了RanGTP依赖性MT稳定,并显著降低纺锤体组装速率。同时耗尽TPX2完全抑制中心体依赖性纺锤体组装。我们的结果表明,Cdk11负责染色体周围RanGTP依赖性MT的稳定,并且这种局部稳定对于正常的纺锤体组装速率和纺锤体功能至关重要。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a7c/2265385/c3d27b5599dc/jcb1800867f01.jpg

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