Host Lipids Associated with Genome Replication in Flock House Virus
Dr. Benjamin Kopek
Positive-strand RNA [(+)RNA] viruses are significant human pathogens. A universal feature of (+)RNA viruses is that they replicate their genomes in association with host intracellular membranes. This association may be a target for broad spectrum antivirals against (+)RNA viruses. The (+)RNA virus we used for our studies is Flock House virus (FHV). FHV is a simple (+)RNA virus with a 4.5 kb bipartite genome that replicates in insect cells; Drosophilia melanogaster cells were used in this study. Previous work by others has shown an involvement of specific lipids, including phosphatidylcholine (PC), in FHV genome replication. To extend this work, we performed genetic knockouts of PC biosynthesis genes using Crispr/Cas9 genome editing. Lipid analysis indicated a reduction in total cellular PC content in these cells. Importantly, qPCR data showed a reduction in FHV RNA replication in cells where PC biosynthesis genes were knocked out. Next, we sought to more specifically examine the role of lipids in FHV genome replication by localizing lipids relative to sites of virus replication. We observed no significant enrichment of phosphatidylethanolamine (PE) or phosphatidylserine (PS) near or at sites of FHV replication. We are currently optimizing studies for imaging PC as well as lipid droplets. These initial studies have provided promising data regarding the role of specific lipids in (+)RNA virus replication.
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