Investigation of the Role of Gle1 in mRNA Export

Gle1 在 mRNA 输出中的作用研究

• 批准号: 6998212
• 负责人: Elizabeth J Tran
• 金额: $ 4.4万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-01-01 至 2007-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6998212
• 关键词: Saccharomyces cerevisiae; affinity chromatography; chemical stability; fluorescent in situ hybridization; fungal genetics; fungal proteins; gel mobility shift assay; gene deletion mutation; gene expression; intracellular transport; mass spectrometry; messenger RNA; nuclear membrane; nuclear transport; open reading frames; postdoctoral investigator; protein localization; protein protein interaction; protein purification; protein structure function; ribonucleoproteins; temperature sensitive mutant; transport proteins

DESCRIPTION (provided by applicant): Export of messenger RNA transcripts (mRNAs) from the nucleus to the cytoplasm is an inherent property of eukaryotic gene expression, however, the mechanism is poorly understood. Recently, the Wente laboratory discovered a physical interaction between the conserved mRNA export factor, Gle1 and the nuclear poly (A)- binding protein, Nab2. The goal of this proposal is to characterize the Gle1: Nab2 complex as a critical, terminal step in mRNA export using the budding yeast S. cerevisiae as a model system. Results gained from this proposal will delineate the first functional role of Gle1 and will be a profound advancement for the field of gene expression. In aim I, will use classic biochemistry coupled with cutting-edge mass spectrometry to determine the nature of the Gle1: Nab2 complex in yeast as well as the stability of this complex in mutant backgrounds. Additionally, I will employ techniques in yeast genetics and cellular microscopy to characterize any newly identified, novel mRNA export factors. In aim II, I will develop an in vitro assembly system to determine if the Gle1: Nab2 complex associates with RNA and if RNA-binding affinity is altered by addition of other members of the Gle1Nab2 complex, this provides a powerful, in vitro complement to aim I.

描述（由申请人提供）：从细胞核到细胞质的使者RNA转录物（mRNA）的导出是真核基因表达的固有特性，但是，该机制知之甚少。最近，温特实验室发现了保守的mRNA输出因子GLE1和核聚（A） - 结合蛋白Nab2之间的物理相互作用。该提案的目的是将GLE1：NAB2复合物表征为使用酿酒酵母S. cerevisiae作为模型系统的关键终端步骤。从该建议中获得的结果将描述GLE1的第一个功能作用，这将是基因表达领域的深刻进步。在AIM I中，将使用经典的生物化学以及尖端的质谱法来确定酵母中GLE1：Nab2复合物的性质以及在突变体背景下该复合物的稳定性。此外，我将在酵母遗传学和细胞显微镜中采用技术来表征任何新发现的新型mRNA输出因子。在AIM II中，我将开发一个体外组装系统，以确定GLE1：NAB2复合物是否与RNA相关联，以及是否通过添加GLE1NAB2复合物的其他成员来改变RNA结合亲和力，这为AIM I提供了强大的体外补体。