A FUNCTIONAL STUDY OF NATIVE AND SYNTHETIC NUCLEI

真核细胞被分成许多膜封闭的细胞， 细胞器 细胞生物学家最感兴趣的是 这些细胞器由此获得独特的组成。 这个目标 建议是理解大分子的选择性运输 进入细胞核。 这种转运通过核孔进行， 包裹着核膜，包括选择性地 入口受到控制。 提出了两种解决该机制的方法 通过孔的运输：1）分析特定的孔蛋白， 63kd糖蛋白，和2）分析核的需要， 运输自己。 使用允许快速体外测定核转运的系统， 我们已经鉴定了孔功能所需的核孔蛋白。 一 对该蛋白进行了分子和功能分析。 蛋白质 将被纯化并产生抗体。有了这些工具， 孔功能的基本问题将被问到，包括如何 蛋白质参与，蛋白质的碳水化合物部分在 功能，以及这种蛋白质是否是结构的组成部分， 原子核 在理解核运输的第二种方法中， 实验提出了确定最小细胞成分 参与运输。 应该可以确定 载体蛋白是核运输所必需的，因为它是输入到 其他细胞器 最后，ATP在识别步骤中的作用 或核运输的易位步骤将被解决。

A eukaryotic cell is compartmentalized into numerous membrane-enclosed organelles. Of fundamental interest to cell biologists are the mechanisms by which these organelles achieve unique compositions. The goal of this proposal is an understanding of the selective transport of macromolecules into the nucleus. Such transport occurs through nuclear pores, which perforate the nuclear envelope and comprise the sites at which selective entry is controlled. Two approaches are proposed to address the mechanism of transport through the pores: 1) analysis of a particular pore protein, a 63 kd glycoprotein, and 2) analysis of the requirements for nuclear transport itself. Using a system which allows the rapid in vitro assay of nuclear transport, we have identified a nuclear pore protein required for pore function. A molecular and functional analysis of this protein is porposed. The protein will be purified and antibodies raised against it. With these tools, fundamental questions of pore function will be asked, including how the protein is involved, the role of the carbohydrate moiety of the protein in function, and whether this protein is integral to the structure of the nucleus. In a second approach to understanding nuclear transport, experiments are proposed to determine the minimal cellular components involved in transport. It should be possible to determine whether a carrier protein is required for nuclear transport, as it is for import into other organelles. Lastly, the role of ATP in either the recognition step or the translocation step of nuclear transport will be addressed.