Investigation of Lipid‐Based Assembly of Viral Particles

Abstract

Sustainment of viral infections from HIV, Ebola and the Human T‐Cell Leukemia Virus (HTLV) requires assembly at the host cell membrane to replicate via budding into new infectious viral particles. The assembly process requires lipid binding by viral matrix proteins as well as hijacking of host cell machinery to complete the budding and replication process. Currently, there is a paucity of information on the molecular architecture of this process including how lipids in the plasma membrane regulate the budding process. To analyze the lipids present when a virus buds from the plasma membrane we have begun a comprehensive and interdisciplinary investigation of the budding process. We have investigated the lipid binding and cellular targeting mechanisms of viral matrix proteins to understand their role in targeting the plasma membrane bud site. Next, we purified virus like particles (VLPs) from a number of infected human host cells to assess their lipid composition. The lipid compositions of these strains were then compared to the lipid composition of the plasma membrane of host cells via lipid mass spectrometry. Lastly, we performed a variety of mutagenic and lipid signaling assays to assess the role of different cofactors in VLP formation. Results elucidate the role of matrix proteins in viral budding and host cell lipid changes responsible for sustaining different types of viral infections.