Translational Regulation by the TNF Alpha AU-rich Element

富含 TNF Alpha AU 元素的翻译调控

• 批准号: 6726750
• 负责人: STUART W PELTZ
• 金额: $ 29.02万
• 依托单位: UNIV OF MED/DENT NJ-R W JOHNSON MED SCH
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-12-01 至 2008-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6726750
• 关键词: RNA interference; Saccharomyces cerevisiae; animal tissue; biological signal transduction; electroporation; fluorescence resonance energy transfer; gene interaction; genetic manipulation; genetic screening; green fluorescent proteins; messenger RNA; nucleic acid denaturation; posttranslational modifications; protein structure function; regulatory gene; tumor necrosis factor alpha

TNFalpha is a pro-inflammatory cytokine produced transiently by several cell types in response to infection or injury. Production of TNFalpha is tightly regulated at the levels of transcription, mRNA decay and translation and aberrant expression of this potent molecule has been linked to autoimmune disorders, rheumatoid arthritis, Crohn's disease and other inflammatory conditions. The translational regulation of TNFalpha is mediated by AU-rich elements (AREs) located in the 3'-UTR of the mRNA. The ARE is involved in translational repression in resting cells as well as in activation of translation under stimulatory conditions. Several ARE-binding proteins have been identified and, of these, TIA-1 and TIAR have been implicated in translational repression. However, the factors involved in up-regulation of TNFalpha translation are not known. A system for studying this phenomenon has been developed in the yeast Saccharomyces cerevisiae. The yeast homologues of the ARE-binding protein Tristetraprolin (TTP) and a yeast homologue of TIA-1/TIAR were found to be key mediators of translation regulation by the ARE. The goals of this proposal are to utilize the yeast system to characterize the mechanism of translational regulation by the TNFalpha ARE, and to perform a genetic screen to identify the trans-acting factors required for accurate regulation. In addition, the role of TTP and its murine homologues (Tis 11 b and Tis 11 d) in modulating TNFalpha translation in mouse macrophages will be investigated. The experiments described here aim to increase our understanding of both general ARE function and regulation of TNFalpha expression.

tnfalpha是一种促炎性细胞因子，该因子响应感染或损伤而瞬时产生。在转录，mRNA衰减和翻译水平上，TNFALPHA的产生与自身免疫性疾病，类风湿关节炎，克罗恩病和其他炎症疾病有关。 TNFALPHA的翻译调节是由位于mRNA的3'-UTR中的Au富元素（ARE）介导的。 IS参与静息细胞中的翻译抑制以及在刺激条件下翻译的激活。已经确定了几种结合蛋白，其中TIA-1和TIAR与翻译抑制有关。但是，尚不清楚tnfalpha翻译上调的因素。研究这种现象的系统是在酿酒酵母中开发的。发现TIA-1/TIAR的酵母菌蛋白三烷酸（TTP）的酵母同源物是由IS进行翻译调节的关键介体。该提案的目标是利用酵母系统来表征TNFALPHA翻译调节的机制，并执行遗传筛选以识别跨性别 准确调节所需的因素。此外，将研究TTP及其鼠同源物（TIS 11 B和TIS 11 D）在调节小鼠巨噬细胞中TNFalpha翻译中的作用。此处描述的实验旨在提高我们对一般的理解是tnfalpha表达的功能和调节。