MINORITY PREDOCTORAL FELLOWSHIP PROGRAM

本提案的总体目标是研究NTF 2介导的Ran输入的机制。 Ran GT3是所有真核细胞中一个重要过程的核心组成部分-核运输。 该实验室和其他实验室的结果表明，小GTdR Ran的核输入由核转运因子2（NTF 2）介导，定义了一种新的Ran输入途径。 在这里，我们将根据我们的Ran导入模型测试三个假设。 使用体外生物化学和透化细胞测定法，我们将研究NTF 2螯合和RanGDP靶向核孔对Ran输入的影响。 此外，我们将使用定量交换测定和新型透化细胞测定来测试Ran从核孔释放所需的RCC 1交换活性的预测。 最后，我们将研究Nup 153在Ran转运中的功能，Nup 153是一种候选NTF 2：RanGDP核孔对接蛋白，被提议在其他转运途径中发挥作用。 这些假设的调查可能会揭示新的信息NTF 2介导的RAN运输所需的分子相互作用，无论是在运输的可溶相和在固定相在核孔。 这些新数据可能为我们进一步理解核运输提供关键信息，核运输是所有真核细胞中生命绝对需要的动态过程。

The overall goal of this proposal is to investigate the mechanism of NTF2-mediated Ran import. The Ran GTPase is a central component of an essential process in all eukaryotic cells -- nuclear transport. Results from this laboratory and others demonstrate that nuclear import of the small GTPase Ran is mediated by Nuclear Transport Factor 2 (NTF2), defining a novel Ran import pathway. Here, we will test three hypotheses based on our model for Ran import. Using in vitro biochemistry and a permeabilized cell assay, we will investigate the effect on Ran import of NTF2 sequestration and targeting of RanGDP to the nuclear pores. Additionally, we will test the prediction that RCC1 exchange activity is required for release of Ran from nuclear pores using quantitative exchange assays and a novel permeabilized cell assay. Finally, we will investigate the function in Ran transport of Nup 153, a candidate NTF2:RanGDP nuclear pore docking protein that is proposed to function in other transport pathways. Investigation of these hypotheses is likely to reveal new information as to the molecular interactions required for NTF2-mediated Ran transport, both in the soluble phase of transport and in the stationary phase at the nuclear pore. These new data may provide information critical to our further understanding of nuclear transport, a dynamic process absolutely required for life in all eukaryotic cells.