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

翻译调控是整个发育过程中调控基因表达的重要机制。然而，翻译激活和抑制的分子机制尚不清楚。来自秀丽隐杆线虫的GLD-1蛋白属于RNA信号转导和激活（STAR）家族的RNA结合蛋白，该家族的成员从酵母到人类都在进化上保守，对许多发育过程都很重要。GLD-1在秀丽隐杆线虫雌雄同体生殖系干细胞发育中起多种作用。其最被研究的功能之一是抑制秀丽隐杆线虫性别决定基因tra-2的翻译。GLD-1通过与tra-2信使RNA （mRNA）中的调节元件TGE结合而起作用。在这个项目中，GLD-1/RNA复合物的三维结构模型将使用核磁共振（NMR）光谱来构建，这将导致对这种蛋白质-RNA相互作用的详细了解。对于核磁共振的结构测定，将采用不同的特定化学标记方法，如对糖基部分进行部分氘化或对RNA内的核碱基进行选择性氟化。

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.