High resolution imaging of the dengue virus replication cycle
Dengue virus (DENV) is a mosquito-borne (+)RNA virus that is responsible for an estimated ~390 million infections and ~25,000 deaths worldwide each year.
Disease symptoms range from mild fever to life-threatening dengue haemorrhagic fever (DHF) and dengue shock syndrome (DSS). Furthermore, the global impact of DENV is expected to increase due to the rapid geographical expansion of the main mosquito vector Aedes aegyptii.
Accordingly, there is an urgent need for the development of safe and effective antiviral therapies to combat DENV infection and associated pathogenic effects. An improved understanding of the molecular details of the dengue virus replication cycle will facilitate the development of such antiviral therapies.
Recently we used high-throughput genome-wide transposon mutagenesis coupled to next-generation sequencing (NGS) to unveil regions of DENV genetic flexibility at high resolution. This revealed remarkable plasticity of discrete regions within the viral NS1 protein and enabled the generation of the first NS1-tagged infectious reporter viruses . NS1 is essential to viral RNA replication and infectious virus production, although exactly how it contributes to these processes remains unclear.
In this project we will use these unique reporter viruses, in combination with new and established high resolution live cell imaging, electron microscopy and super-resolution imaging techniques [2-3], to investigate the localisation and traffic of NS1 protein during an infectious dengue virus replication cycle. This will reveal new details about NS1-host interactions and may unveil novel targets for future antiviral drug development.
- Eyre NS, Johnson SM, Eltahla AA, et al. (2017) Genome-Wide Mutagenesis of Dengue Virus Reveals Plasticity of the NS1 Protein and Enables Generation of Infectious Tagged Reporter Viruses.
- Eyre NS, Hampton-Smith RJ, Aloia AL, et al. (2016) Phosphorylation of NS5A Serine-235 is essential to hepatitis C virus RNA replication and normal replication compartment formation.
- Eyre NS, Fiches GN, Aloia AL, et al. (2014) Dynamic imaging of the hepatitis C virus NS5A protein during a productive infection.