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

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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.