Poster Presentation 28th Annual Lorne Proteomics Symposium 2023

The unfolded protein response and dynamic host subcellular proteome in response to H1N1 and H3N2 influenza virus infection (#141)

Kyle Macauslane 1 , Cassandra Pegg 1 , Kirsty Short 1 , Ben Schulz 1
  1. School of Chemistry and Molecular Biosciences, University of Queensland, Brisbane, QLD, Australia

Influenza viruses, particularly influenza A virus (IAV), are responsible for seasonal and pandemic outbreaks of Influenza, a respiratory disease which heavily burdens the elderly population. Two subtypes of IAVs are commonly transmitted amongst humans, the H1N1 and H3N2 subtypes. Seasonal outbreaks that are dominated by the H3N2 virus tend to be more severe. Due to both the emergence of antiviral resistance, and the lack of a universal vaccine for influenza viruses, developing our understanding of viral-host protein interactions remains essential. Although global proteomic changes in influenza infection have previously been investigated, little attention has been given to characterising these changes at a subcellular level or with consideration to protein post translational modifications such as glycosylation. We used a subcellular fractionation protocol to isolate cellular compartments relevant to influenza virology: the nucleus, cytoplasm, organelle, and secreted proteins. We developed an LC-MS/MS approach combining tandem mass tag (TMT) labelling and hydrophilic liquid interaction chromatography (HILIC) enrichment of glycopeptides. Using this approach, we are able to quantify and compare protein and glycoprotein abundances in each fraction throughout a time-course infection of human A549 cells with two influenza A strains: the H1N1 virus strain A/Puerto Rico/8/1934, and the H3N2 strain A/X-31. Here, we demonstrate the dynamic subcellular host response to IAV infection and describe strain-specific responses observed in relation to the unfolded protein response (UPR).