Dissecting the role of the nuclear basket in regulating mRNA metabolism

调节其基因的表达对任何活的有机体来说都是一项基本任务。在真核细胞中，存储遗传信息的部位，即细胞核，以及将信息转化为蛋白质的部位，即细胞质，被一层不被大分子渗透的双层胆脂膜隔开。在储存DNA的细胞核中，遗传信息首先被复制到称为信使分子的信使分子中，信使分子必须离开细胞核，到达细胞质才能翻译成蛋白质。到达蛋白质合成部位的唯一途径是通过核孔复合体(NPC)，这是一种嵌入核膜的大分子复合体。我们研究计划的长期目标是了解与mRNA结合的不同因素和组成NPC的因素之间的相互作用如何控制mRNAs从细胞核到细胞质的有效和选择性输出。为了进一步了解这一重要过程，我们将使用不同的生物物理、生化和遗传工具。首先，我们将使用最近发展的超分辨率显微镜技术，允许实时和纳米分辨率跟踪单个mRNA分子，以确定核篮子是如何调节mRNPs进入鼻咽癌的。核篮子是mRNAs到达核外围时遇到的第一个结构。此外，利用基因组和蛋白质组分析，我们将研究当细胞暴露在压力下时，细胞如何诱导鼻咽癌成分的变化来改变基因表达程序。通过完成这个项目，我们将揭示这一基本细胞过程背后的重要机制细节，并建立相关的生物物理工具，以单分子分辨率研究不同的细胞过程。

Regulating the expression of its genes is an essential task for any living organism. In eukaryotic cells, the site where the genetic information is stored, the nucleus, and the site where the information is translated into protein, the cytoplasm, is separated by a double bilipidic membrane, impermeable to large molecules. In the nucleus, the organelle where DNA is stored, the genetic information is first copied into a messenger molecule, called mRNA, which has to exit the nucleus and reach the cytoplasm to get translated into proteins. The only path to reach the site of protein synthesis is through the nuclear pore complex (NPC), a large macromolecular complex embedded in the nuclear membrane. The long-term objective of our research program is to understand how the interplay between different factors that bind to the mRNA and factors composing the NPC control efficient and selective export of mRNAs from the nucleus to the cytoplasm. To gain further insight into this essential process, we will use different biophysical, biochemical and genetic tools. First, we will use recently developed super resolution microscopy techniques that allow to follow individual mRNA molecules in real time and in nanometer resolution to determine how nuclear basket, the first structure mRNAs encounter when reaching the nuclear periphery, regulates the access of mRNPs to the NPC. Furthermore, using genomic and proteomic assays, we will investigate how cells induce changes in the composition of the NPC to change gene expression programs when cells are exposed to stress. By completing this project we will reveal important mechanistic details that underlie this essential cellular process and establish biophysical tools relevant to study different cellular processes in single molecule resolution.