Lightning Oral Talk 25th Annual Lorne Proteomics Symposium 2020

Studying uncharacterised non-structural proteins of Rabbit haemorrhagic disease virus  (#63)

Elena Smertina 1 2 , Robyn Hall 2 , Tanja Strive 2 3 , Michael Frese 1 2
  1. Faculty of Science and Technology, University of Canberra , Canberra , ACT, Australia
  2. Health & Biosecurity , CSIRO, Canberra, ACT, Australia
  3. Invasive animals cooperative research centre, University of Canberra , Canberra, ACT, Australia

The Rabbit haemorrhagic disease virus (RHDV) is a highly virulent virus that cannot be grown in cell culture. The functions of several non-structural proteins of RHDV (p16, p23 and p29) are still unknown. It is generally assumed that these proteins counteract the host's antiviral defense, similar to the certain norovirus proteins, e.g., p48. Norovirus protein p48 affects the organisation of intracellular membranes and thus inhibits secretory pathways. The functions of RHDV p16, p23 and p29 could be deduced through the identification of their cellular and viral interaction partners. In addition, changes to whole cell proteome upon the expression of the proteins of interest can be tracked. To identify protein interaction partners, we utilised a pull-down assay with FLAG-tagged viral proteins and anti-FLAG resin. Several possible cellular interaction partners were identified after co-immunoprecipitation and quantified using mass spectrometry-based SILAC (stable isotope labelling in cell culture) approach. Preliminary results indicate that proteins p23 and p29 may take part in the disruption of intracellular membranes with subsequent formation of vesicular compartments for viral replication and interact with cellular stress-induced proteins.