Investigation of the Role of Gle1 in mRNA Export

Gle1 在 mRNA 输出中的作用研究

• 批准号: 7167434
• 负责人: Elizabeth J Tran
• 金额: $ 4.88万
• 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/7167434
• 关键词: Affinity; Affinity Chromatography; Bacteria; Biochemistry; Biological Assay; Biological Models; Cell Nucleus; Complement; Complex; Coupled; Coupling; Cytoplasm; Gene Expression; Genes; Goals; In Vitro; Investigation; Laboratories; Life; Mass Spectrum Analysis; Messenger RNA; Microscopy; Nature; Nuclear; Nuclear Pore; Poly(A)+ RNA; Poly(A)-Binding Proteins; Process; Property; Proteins; RNA; RNA Binding; Ribonucleoproteins; Role; Saccharomyces cerevisiae; Saccharomycetales; System; Techniques; Transcript; Yeasts; base; functional group; mRNA Export; member; mutant; novel; trafficking; yeast genetics

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 stabiility 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.

将信使 RNA 转录本 (mRNA) 从细胞核输出到细胞质是细胞的固有特性 然而，对于真核基因表达的机制却知之甚少。近日，文特实验室 发现了保守的 mRNA 输出因子 Gle1 和核聚 (A)- 之间的物理相互作用 结合蛋白，Nab2。该提案的目标是将 Gle1:Nab2 复合物描述为关键的、 使用芽殖酵母酿酒酵母作为模型系统进行 mRNA 输出的最终步骤。取得的成果 该提案中的内容将描绘 Gle1 的第一个功能角色，并将是对 基因表达领域。在目标 I 中，将使用经典生物化学与尖端质谱法相结合 确定酵母中 Gle1:Nab2 复合物的性质以及该复合物在突变体中的稳定性 背景。此外，我将采用酵母遗传学和细胞显微镜技术来表征 任何新发现的新型 mRNA 输出因子。在目标 II 中，我将开发一种体外组装系统 确定 Gle1:Nab2 复合物是否与 RNA 结合，以及 RNA 结合亲和力是否因添加而改变 Gle1Nab2 复合物的其他成员，这为 Target I 提供了强大的体外补充。