Replication and Assembly of Influenza virus

The theme of the proposed research is our understanding of the replication and assembly of Influenza virus, from a structural imaging perspective. The transcription and replication of the influenza virus genome occurs in the nucleus and the structure of the isolated polymerase reveals at a molecular level key steps in transcription and replication. The genome of Influenza virus is composed of 8 RNA segments each bound to the viral RNA polymerase and multiple copies of N protein to form ribonucleoprotein complexes (vRNPs). Newly replicated vRNPs are transported out of the nucleus, where they are trafficked to the cell membrane and assemble to form new viruses budding out from the cell membrane. How the production and assembly of RNPs is coordinated in the nucleus and the details of how the vRNPs are transported out of the nucleus to the cell membrane is unknown. By integrating structural, biochemical and cellular imaging, I aim to understand with atomic-level detail, vRNP production, nuclear export and viral assembly. This will be done using crystallography and single particle cryo-EM, combined with high resolution cellular EM and X-ray tomography, correlated with cryo- fluorescence imaging of infected cells to understand these key stages in the viral cycle.