Structure and Mechanism of the Ire 1 Riboedonuclease

Ire 1 核糖核酸酶的结构和机制

• 批准号: 7113281
• 负责人: BRIAN R BOWMAN
• 金额: $ 5.04万
• 依托单位: HARVARD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-04 至 2009-04-03
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7113281
• 关键词: JUN kinase; RNA splicing; X ray crystallography; adenosine triphosphate; calorimetry; combinatorial chemistry; endoplasmic reticulum; endoribonucleases; enzyme activity; enzyme complex; enzyme mechanism; enzyme structure; high throughput technology; immunoprecipitation; inhibitor /antagonist; insulin receptor; nucleic acid structure; nucleoside analog; postdoctoral investigator; protein folding; protein structure function; tissue /cell culture; western blottings

DESCRIPTION (provided by applicant): Eukaryotic cells have evolved a novel endoplasmic reticulum (ER) to nucleus signaling pathway, the unfolded protein response (UPR) to respond to conditions of ER stress. UPR induction is initiated by the activation of Ire1, an ER-resident kinase and endoribonuclease. UPR disregulation is implicated in viral infection, Alzheimer's disease, and with insulin resistance and type 2 diabetes. This proposal describes experiments designed to understand the molecular mechanisms of Ire1 signal generation in the UPR signaling pathway using small molecule inhibitors. A cell-based assay in combination with high throughput technologies will be used to screen libraries of thousands of small molecules for Ire1 inhibition. These inhibitors will be used as tools to probe their effects on UPR signal transduction and downstream signaling cascades. In vitro biochemical experiments with purified Ire1, using a combination of biochemistry and x-ray crystallography, will be initiated to understand the structure/function relationships of Ire signaling as well as the mode of inhibition of these small molecules at atomic resolution. Understanding the mechanisms of Ire1 signaling will facilitate the modulation of UPR signaling thus aiding in the treatment of UPR related diseases.

描述（由申请人提供）：真核细胞已经进化出一种新的内质网（ER）到细胞核信号传导途径，即未折叠蛋白反应（UPR）以对ER应激条件做出反应。 UPR 诱导是由 Ire1（一种 ER 驻留激酶和核糖核酸内切酶）的激活启动的。 UPR 失调与病毒感染、阿尔茨海默病、胰岛素抵抗和 2 型糖尿病有关。该提案描述了旨在了解使用小分子抑制剂在 UPR 信号通路中 Ire1 信号生成的分子机制的实验。基于细胞的测定与高通量技术相结合将用于筛选数千个小分子库以抑制 Ire1。这些抑制剂将用作探讨其对 UPR 信号转导和下游信号级联的影响的工具。将结合生物化学和 X 射线晶体学，对纯化的 Ire1 进行体外生化实验，以了解 Ire 信号传导的结构/功能关系以及这些小分子在原子分辨率下的抑制模式。了解 Ire1 信号传导机制将有助于 UPR 信号传导的调节，从而有助于治疗 UPR 相关疾病。