Structure and activity of a cellular IRES element

细胞 IRES 元件的结构和活性

• 批准号: 7389473
• 负责人: Andrew Mehle
• 金额: $ 5.04万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-07 至 2009-04-06
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7389473
• 关键词: Biological Assay; Chemicals; Complex; Crystallization; Elements; Genetic Translation; Goals; Indium; Internal Ribosome Entry Site; Link; Messenger RNA; Peptide Initiation Factors; Play; Production; Proteins; RNA; Reporter; Resolution; Ribosomes; Role; Site; Structure; Trans-Activators; Translation Initiation; Viral; Work; tumor progression

Translation of mRNA is tightly regulated, requiring the ordered assembly of initiation factors and ribosomal subunits. Certain viral and cellular mRNAs circumvent traditional requirements for translation initiation by utilizing structured RNA elements termed internal ribosome entry sites (IRESs) to direct ribosomes to the translational start site. IRES structure is intimately linked to function, but the three-dimensional organization of cellular IRESs is poorly understood. This proposal aims to obtain a crystallographic structure of the IRES element in Cyr61 mRNA in complex with proteins required for its function, providing the first high-resolution structure of a cellular IRES element. Cyr61 is an angiogenic inducer suggested to play a critical role in tumor progression. Dicistronic reporter assays will define a minimal functional IRES and trans-acting factors required for activity will be identified. Chemical and enzymatic probing will determine the secondary structure of the Cyr61 IRES and identify structural elements amenable to crystallization. Results from this work will yield a detailed mechanistic understanding of the Cyr61 IRES and may elucidate mechanisms of cellular IRES activity, possibly identifying strategies to therapeutically manipulate Cyr61 production.

mRNA的翻译受到严格调节，需要有序的起始因子和核糖体的组装 亚基。某些病毒和细胞mRNA避免了传统要求对翻译启动的要求 利用称为内部核糖体进入位点（IRESS）的结构化RNA元素将核糖体直接直接 翻译起始站点。 IRES结构与功能密切相关，但三维组织 细胞IRESS的理解很少。该建议旨在获得IRES的晶体学结构 CYR61 mRNA中的元素与其功能所需的蛋白质复合物中的元素，提供了第一个高分辨率 细胞IRES元件的结构。 CYR61是一种血管生成诱导剂，建议在 肿瘤进展。 DICISTRONIC RETORTER分析将定义最小功能IRE和反式因素 将确定活动所需的。化学和酶探测将确定次要 CYR61 IRES的结构，并识别可符合结晶的结构元素。结果 工作将对CYR61 IRES产生详细的机械理解，并可能阐明 细胞IRES活性，可能识别治疗策略对CYR61生产进行操纵。

项目成果

An inhibitory activity in human cells restricts the function of an avian-like influenza virus polymerase.

• DOI: 10.1016/j.chom.2008.06.007
• 发表时间: 2008-08-14
• 期刊: Cell host & microbe
• 影响因子: 30.3
• 作者: Mehle A;Doudna JA
• 通讯作者: Doudna JA