Structure determination of the TGE element of tra-2 messenger RNA in complex with the posttranscriptional regulatory protein GLD-1 by nuclear magnetic resonance spectroscopy

核磁共振波谱法测定 tra-2 信使 RNA 与转录后调节蛋白 GLD-1 复合物的 TGE 元件的结构

• 批准号: 21052241
• 负责人: Dr. Christine Beuck
• 金额: --
• 依托单位: Skaggs Institute for Chemical Biology
• 依托单位国家: 德国
• 项目类别: Research Fellowships
• 财政年份: 2005
• 资助国家: 德国
• 起止时间: 2004-12-31 至 2008-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/21052241?language=en
• 关键词: Structure; determination; TGE; element; tra

Translational control is an essential mechanism of controlling gene expression throughout development. Yet the molecular mechanism of translational activation and repression are poorly understood. The protein GLD-1 from the nematode worm Caenorhabditis elegans belongs to the signal transduction and activation of RNA (STAR) family of RNA-binding proteins, whose members are evolutionarily conserved from yeast to humans and are important for a number of developmental processes. GLD-1 plays multiple roles in the germ line stem cell development of C. elegans hermaphrodites. One of its best studied functions is the inhibition of translation of the C. elegans sex-determining gene tra-2. GLD-1 acts through binding to regulatory elements, called TGE, within the tra-2 messenger RNA (mRNA). In this project, a model of the three-dimensional structure of the GLD-1/RNA complex will be constructed using nuclear magnetic resonance (NMR) spectroscopy which should lead to detailed understanding of this protein-RNA interaction. For the structure determination by NMR, different specific chemical labeling methods, like partial deuteration of the rebose moieties or selective fluorination of the nucleobases within the RNA, will be applied.

翻译调控是整个发育过程中控制基因表达的一种重要机制。然而，翻译激活和抑制的分子机制却知之甚少。秀丽线虫的GLD-1蛋白属于RNA结合蛋白信号转导和激活(STAR)家族，其成员在从酵母到人类的进化过程中都是保守的，在许多发育过程中起重要作用。GLD-1在线虫两性生殖系干细胞发育中发挥多种作用。它被研究得最好的功能之一是抑制线虫性别决定基因tra-2的翻译。GLD-1通过与tra-2信使RNA(MRNA)中称为TGE的调节元件结合来发挥作用。在这个项目中，将使用核磁共振(NMR)光谱建立GLD-1/RNA复合体的三维结构模型，这将导致对这种蛋白质-RNA相互作用的详细了解。对于核磁共振结构的测定，将使用不同的特定化学标记方法，如核糖体部分的部分氢化或核糖核酸碱基的选择性氟化。