Dissecting the role of the nuclear basket in regulating mRNA metabolism

剖析核篮在调节 mRNA 代谢中的作用

• 批准号: RGPIN-2015-05922
• 负责人: Zenklusen, Daniel
• 金额: $ 2.77万
• 依托单位: Université de Montréal
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2016
• 资助国家: 加拿大
• 起止时间: 2016-01-01 至 2017-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=613826
• 关键词: Dissecting; role; nuclear; basket; regulating

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.

调节其基因的表达是任何生物体的必不可少的任务。在真核细胞中，遗传信息存储的位置，核和信息转化为蛋白质，细胞质的位点，由双重双膜膜分离，不可能对大分子进行渗透。在原子核中，储存DNA的细胞器，首先将遗传信息复制到一个称为mRNA的信元分子中，该分子必须退出细胞核并到达细胞质以转化为蛋白质。到达蛋白质合成部位的唯一途径是通过核孔复合物（NPC），这是一种嵌入核膜中的大型大分子复合物。我们的研究计划的长期目标是了解如何与MRNA结合的不同因素与组成NPC控制有效和选择性导出的因素之间的相互作用，将mRNA从细胞核中从细胞核到细胞质。为了进一步了解这一基本过程，我们将使用不同的生物物理，生化和遗传工具。首先，我们将使用最近开发的超级分辨率显微镜技术，该技术允许实时和纳米分辨率遵循单个mRNA分子，以确定核篮（核篮）在到达核外围时的第一个结构mRNA遇到的核篮如何调节mRNPS对NPC的访问。此外，使用基因组和蛋白质组学测定法，我们将研究细胞如何在细胞暴露于胁迫的情况下改变NPC的组成改变基因表达程序。通过完成该项目，我们将揭示重要的机械细节，这些细节是该基本细胞过程的基础，并建立了与研究单分子分辨率不同细胞过程相关的生物物理工具。