Regulation of microtubule assembly and organization in mitosis by the AAA plus ATPase pontin
Regulation of microtubule assembly and organization in mitosis by the AAA plus ATPase pontin
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DOI：
10.1091/mbc.e07-11-1202
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发表时间：
2008-07-01
影响因子：
3.3
通讯作者：
Zheng, Yixian
中科院分区：
文献类型：
--
作者：
Ducat, Daniel;Kawaguchi, Shin-ichi;Zheng, Yixian
为了鉴定在有丝分裂中对微管组装重要的新蛋白，我们使用了基于中心体的互补测定来富集具有有丝分裂功能的蛋白。基于RNA干扰（RNAi）的这些蛋白质的筛选使我们能够发现13种新的有丝分裂调节因子。我们对其中一种蛋白质Pontin进行了深入分析，已知Pontin在间期具有多种功能，包括染色质重塑，DNA修复和转录。我们发现，减少Pontin的RNAi导致缺陷的纺锤体组装在果蝇S2细胞和几种哺乳动物组织培养细胞系。对非洲爪蟾卵提取物中桥蛋白的进一步表征表明桥蛋白与γ微管蛋白环复合物（γ-TuRC）相互作用。由于耗尽的Pontin导致缺陷的装配和组织的微管阵列在鸡蛋提取物中，我们的研究表明，Pontin有一个有丝分裂特异性的功能，在调节微管组装。
To identify novel proteins important for microtubule assembly in mitosis, we have used a centrosome-based complementation assay to enrich for proteins with mitotic functions. An RNA interference (RNAi)-based screen of these proteins allowed us to uncover 13 novel mitotic regulators. We carried out in-depth analyses of one of these proteins, Pontin, which is known to have several functions in interphase, including chromatin remodeling, DNA repair, and transcription. We show that reduction of Pontin by RNAi resulted in defects in spindle assembly in Drosophila S2 cells and in several mammalian tissue culture cell lines. Further characterization of Pontin in Xenopus egg extracts demonstrates that Pontin interacts with the gamma tubulin ring complex (gamma-TuRC). Because depletion of Pontin leads to defects in the assembly and organization of microtubule arrays in egg extracts, our studies suggest that Pontin has a mitosis-specific function in regulating microtubule assembly.