Investigating NP-PB2 interaction as a potential antiviral target to inhibit influenza replication

Presenter Information(s)

Caroline Vanegas

Abstract or Description

Spillover of highly pathogenic avian influenza H5N1 in mammals threatens emergence of influenza virus with pandemic potential. Antivirals are the first line of defense to slow the spread of novel viruses, but overuse selects resistance. This fuels the need to identify new targets for antiviral development. The influenza viral ribonucleoprotein complex is central to the viral life cycle and harbors conserved regions for targeting. We previously identified amino acids within the body domain of nucleoprotein (NP) essential for influenza RNA synthesis and interaction with polymerase basic 2 (PB2). Disruption of NP-PB2 interaction is a viable target for development of new antivirals against influenza. We engineered single-integration lentiviruses and transduced human embryonic lung cells (A549) to integrate DNA encoding NP peptide encompassing amino acids essential for PB2 interaction or control NPbd3 peptide, with 5 amino acid substitutions that eliminate PB2 interaction to determine if NP peptide functions to inhibit NP-PB2 interaction. Transduced cells express GFP and GFP-NP peptide while remaining healthy and viable through multiple passages. Cells were infected with influenza A virus (H3N2) at both low or high virus concentration and cytopathic effect (CPE) was observed post infection, comparing to mock-infected control. Infected cells expressing GFP-NP peptide exhibit less CPE and less viral mRNA than control, demonstrating NP peptide antiviral function. This research establishes disruption of NP-PB2 interaction as a viable target for development of needed new antivirals to protect against influenza. 

Mentor

Dr. Laura Newcomb