Structure-guided approach to assay development for inhibiting Dengue virus replication

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Dengue virus, transmitted by mosquitos in Africa, Asia and central America, infects ~390 million people and kills ~20,00 people, including children, annually, mostly in tropical climates. With no licensed drugs and a vaccine recommended to people only after a previous Dengue infection, new anti-viral therapeutic strategies are needed. A structure-guided approach accelerates drug discovery but requires information about the structure and organization of protein targets. The architecture of the Dengue virus RNA replication complex remains unknown, despite structures of 5 out of its 7 protein subunits being available. Equivalent complexes are targeted in other anti-viral treatments, such as against Hepatitis C (a close relative of Dengue), HIV, SARS-CoV-2 and the Herpes Simplex type 1 virus. However, the assembly of the Dengue virus replication complex remains poorly characterised. Here, a rigorous structural and biophysical study of the two enzymes, non-structural proteins NS3 and NS5, at the heart of the Dengue viral replication complex will enable the development of new high-throughput assays for in vitro screening, while also supporting future in silico drug discovery efforts. This project will involve collecting electron microscopy and small angle X-ray scattering (SAXS) data to build a model of the Dengue replication complex using previous X-ray structures of the individual proteins. Subsequently, new high-throughput assays will be developed to screen small molecules that inhibit the assembly and enzyme activities of this complex.