Delving deeper into the biomechanics of disease
Micro-RNAs (miRNAs) mature in two phases along a pathway involving Drosha-DGCR8 and Dicer-PACT-TRBP processing complexes. These ribonucleases are recruited by multi-domain RNA-binding accessory proteins that interact with structural features specific to immature miRNAs. the 'Biogenesis of oncogenic microRNAs: from the structure of the microRNA processing complexes to the inhibition of the maturation of human oncogenes' (oncoMiRNA-biogenesis) project is working on a multidisciplinary analysis of biomolecular interaction networks involved in the production processes of human miRNA. Aiming to clone, express and screen soluble domains taken from modular processing proteins, project partners will also use nuclear magnetic resonance (NMR) methods to assess protein and RNA folds, and protein-RNA interaction sites. Experiments focus on defining the role of biogenesis proteins, describing RNA recognition mechanisms and arriving at the first atomic resolution image of miRNA processing complexes. Such an approach is expected to lay the foundation for the design and testing of inhibitors of miRNA biogenesis with a view to uncovering the means of probing miRNA function or potential new cancer therapies. To date, the EU-funded project has identified potential domains constructs from multi-domain proteins involved in miRNA production and provided an initial design of constructs from miRNA biogenesis accessory proteins. Partners have also performed automated cloning and expression testing, achieved biophysical and NMR spectroscopy analysis of features of miRNA biogenesis processes, and optimised miRNA in vitro production. other project accomplishments include analysis of certain inter- and intra-molecular interactions, producing mg quantities of RNA for NMR analysis and developing isotope-labelling strategies for NMR-based analysis of biomolecular interfaces. the oncoMiRNA-biogenesis partners expect that ongoing work will result in achieving structural analysis of miRNA biogenesis machinery and mapping protein-protein interactions and protein-RNA interactions by NMR spectroscopy and other bio techniques. They also aim to develop a high-resolution model for describing critical biomolecular interfaces and interactions playing a role in miRNA biogenesis.
