Poster Presentation 12th Australasian Virology Society Meeting 2024

Mapping the Dengue Virus NS1 Protein Microenvironment in Infected Cells (#141)

Siena Centofanti 1 , Alex Colella 2 , Nusha Chegeni 2 , Kamelya Aliakbari 2 , Gustavo Bracho 3 , Jillian M Carr 1 , Tim Chataway 2 , Nicholas S Eyre 1
  1. Flinders Health and Medical Research Institute, Flinders University, Adelaide, South Australia, Australia
  2. Flinders Proteomics Facility, Flinders University, Adelaide, South Australia, Australia
  3. CellScreen SA, Flinders Centre for Innovation in Cancer, Adelaide, South Australia, Australia

Dengue virus (DENV) is a rapidly-spreading mosquito-borne (+)RNA virus that is currently endemic in over 100 countries and is responsible for a major public health burden. Despite this, there are no approved antiviral therapeutics available. The DENV non-structural protein 1 (NS1) plays critical roles in viral RNA replication, infectious virus particle production and viral pathogenesis and has emerged as a major target in the development of vaccines and antivirals. Towards the identification of DENV non-structural protein 1 (NS1)-host factor interactions that can serve as antiviral drug targets, we have mapped the proteomic composition the NS1 microenvironment in live infected cells using an APEX2 proximity labelling-coupled quantitative proteomics approach in conjunction with an APEX2-tagged reporter virus (DENV2-NS1-APEX2). An siRNA screen targeting the top 50 identified NS1-proximal host factors was performed to identify a panel of host factors that are required for DENV infection. It is hoped that the ongoing characterisation of a selection these validated hits using CRISPR/Cas9-mediated host gene knockout, protein-protein interaction assays and high-resolution confocal imaging analysis will reveal novel DENV NS1-host protein interactions that are essential to the viral replication cycle.