The Nuclear Pore Complex: Interphase and Mitotic Function

描述（由申请人提供）：真核细胞的核区室提供了一个关键的专门微环境，并允许对重要的细胞过程进行空间和时间控制。分子在细胞核和细胞质之间的选择性流动是这种亚细胞分裂的基础，并且仅通过称为核孔复合物的大分子结构发生。细胞的许多结构特征的重塑是细胞分裂的先决条件。当细胞进入有丝分裂时，核孔复合体和包围细胞核的膜都被分解，与染色体浓缩和微管重排相一致。事实上，核孔的组成部分在协调重塑事件中起着关键作用。孔蛋白Nup153将被用来作为一个深入了解孔如何有助于细胞内运输和细胞分裂的机制。具体而言，在目标1中，将分析Nup153的2个关键特征，RNA结合和转录依赖性迁移率，以更好地理解RNA货物和孔复合物之间的界面和机制联系。第二个目标集中在Nup153和Nup358中发现的锌指模块的伴侣蛋白在COPI膜重塑机器的有丝分裂募集方面的作用。在第三个目标中，该锌指模块和COPI之间的连接将被靶向破坏，以破译它如何整合到体细胞的细胞周期中。最后，核膜重塑机制中其他参与者的表征将在第四个目标中通过检查主要候选蛋白以及通过相互作用网络分析鉴定的蛋白质来启动。这些研究将阐明基因在间期的表达和有丝分裂的精确分裂的基本方面。收集的信息将影响实际应用，例如优化外源基因的表达，以及理解并最终阻止癌细胞中发现的生长失调。

DESCRIPTION (provided by applicant): The nuclear compartment of eukaryotic cells provides a critical specialized microenvironment and allows for spatial and temporal control over important cellular processes. Selective flow of molecules between the nucleus and cytoplasm underlies this subcellular division and takes place exclusively through macromolecular structures termed nuclear pore complexes. Remodeling of many architectural features of the cell is a prerequisite for cell division. Both the nuclear pore complex and the membranes that enclose the nucleus are disassembled as the cell enters mitosis, in concert with chromosomal condensation and microtubule rearrangements. Indeed, components of the nuclear pore play critical roles in orchestrating remodeling events. The pore protein Nup153 will be used as an inroad into understanding how the pore contributes mechanistically to both intracellular trafficking and cell division. Specifically, in Aim 1, 2 key features of Nup153, RNA binding and transcription-dependent mobility, will be analyzed in order to better understand the interface and mechanistic links between RNA cargo and the pore complex. The second Aim focuses on the role of partner proteins of the zinc finger module found in Nup153 and Nup358 with respect to mitotic recruitment of COPI membrane remodeling machinery. In the third Aim, the connection between this zinc finger module and COPI will be targeted for disruption in order to decipher how it is integrated into the cell cycle of somatic cells. Finally, characterization of other players in the nuclear membrane remodeling machinery will be launched in the fourth Aim by examining both prime candidate proteins as well as proteins identified through analysis of interaction networks. These studies will elucidate fundamental aspects of gene expression at interphase and accurate division at mitosis. The information gathered will impact practical applications, such as optimizing expression of exogenously introduced genes as well as understanding -and ultimately controlling-growth deregulation found in cancerous cells.