FUNCTIONAL STUDY OF NATIVE AND SYNTHETIC NUCLEI

核孔复合体，一个1亿吨的复合体，由 100个蛋白质在多个副本中，是通过它进口的结构， 大量不同蛋白质和RNA底物的输出和循环 发生。该工艺适用于大于~50 kDa的衬底。这项建议 与脊椎动物鼻咽癌的结构和功能有关。第一个目标是寻求 为了剖析两个人大代表的分子相互作用和功能， Nup98和Nup153。这两种蛋白都定位于鼻咽癌的核表面； 有证据表明，Nup98与出口有关，Nup153与进口有关。网站的位置 与靶向分子和其他NPC成员的联系将被绘制成地图。 第二个目的是利用环状薄板，即自发形成的结构 在卵子细胞质中，类似于NPC，但更容易操作。我们的目标是 梳理这个复合体以确定已知的鼻咽癌的分子相互作用 蛋白质和识别新的蛋白质。环状薄片的形成将 也可以在体外进行研究，这也将提供结构信息。在……里面 最终目标是，P.I.将利用一种新的检测方法，使蛋白质结合到 标记配体，标记的分子被添加到环状薄片中 构成反应，并对涉及的标记蛋白进行鉴定。这个化验结果会 用于鉴定新的NPC蛋白，或测试与已知蛋白质的相互作用 全国人大选民。

The nuclear pore complex, a 100-megadalton complex comprised of 100 proteins in multiple copies, is the structure through which the import, export and cycling of a large number of diverse protein and RNA substrates occur. The process is regulated for substrates above~50 kDa. This proposal concerns the structure and function of the vertebrate NPC. The first aim seeks to dissect the molecular interactions and functions of two members of the NPC, Nup98 and Nup153. Both proteins are localized to the nuclear face of the NPC; evidence has linked Nup98 with export and Nup153 with import. The sites of contact with targeting molecules and other members of the NPC will be mapped. The second aim utilizes annulate lamellae, structures that form spontaneously in egg cytoplasm that resemble NPCs but are easier to manipulate. The goal is to tease apart this complex to identify molecular interactions of known NPC proteins and to identify new ones. The formation of the annulate lamellae will also be studied in vitro, which would also provide structural information. In the final aim, the P.I. will utilize a novel assay in which proteins bound to ligands are tagged, the tagged molecules are added to a annulate lamellae constitution reaction, and involved tagged proteins identified. This assay will be used to identify novel NPC proteins, or to test for interactions with known NPC constituents.