The Nuclear Pore Complex: Interphase and Mitotic Function

描述(由申请人提供)：真核细胞的核隔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，并且完全通过被称为核孔复合体的大分子结构发生。细胞的许多结构特征的重塑是细胞分裂的先决条件。随着细胞进入有丝分裂，伴随着染色体凝聚和微管重排，核孔复合体和包裹细胞核的膜都被分解。事实上，核孔的成分在协调重塑事件中发挥着关键作用。毛孔蛋白Nup153将被用作了解毛孔如何对细胞内运输和细胞分裂做出机械贡献的途径。具体地说，在目标1中，将分析Nup153的2个关键特征，即RNA结合和依赖转录的迁移性，以更好地了解RNA货物和孔复合体之间的界面和机制联系。第二个目的是研究在Nup153和Nup358中发现的锌指模块的伙伴蛋白在COPI膜重塑机制的有丝分裂招募中的作用。在第三个目标中，这个锌指模块和COPI之间的联系将被破坏，以破译它是如何整合到体细胞的细胞周期中的。最后，在第四个目标中，将通过研究主要候选蛋白质以及通过分析相互作用网络确定的蛋白质来表征核膜重塑机制中的其他参与者。这些研究将阐明间期基因表达的基本方面和有丝分裂时的准确分裂。收集到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-并最终控制-癌细胞中发现的生长放松调控。

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.