A FUNCTIONAL STUDY OF NATIVE AND SYNTHETIC NUCLEI

真核细胞被分割成许多被包裹的膜。 细胞器。细胞生物学家最感兴趣的是这种机制 这些细胞器通过这些细胞获得独特的成分。这样做的目的是 建议是对大分子选择性运输的理解 进入原子核。这种运输是通过核孔进行的，核孔 使核膜穿孔并包括选择性地 进入受到控制。提出了两种方法来解决这一机制 通过毛孔的运输：1)特定毛孔蛋白的分析， 一种63kd的糖蛋白，以及2)对核的需求分析 交通本身。 使用允许对核运输进行快速体外检测的系统， 我们已经确定了一种毛孔功能所需的核毛孔蛋白。一个 对该蛋白进行了分子和功能分析。这种蛋白质 将被提纯并产生抗体。使用这些工具， 我们将会问到毛孔功能的基本问题，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所必需的 原子核。在理解核运输的第二种方法中， 建议进行实验以确定最小的细胞成分 涉足运输行业。应该可以确定一个 载体蛋白是核运输所必需的，因为它是进口到 其他细胞器。最后，ATP在识别步骤中的作用 否则，核运输的转移步骤将得到解决。

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.