Several members of the phospholipase family have been reported to be involved in hepatitis C virus (HCV) replication. Here, we identified another phospholipase, phosphatidylserine-specific phospholipase A1 (PLA1A), as a host factor involved in HCV assembly. PLA1A was upregulated by HCV infection, and PLA1A knockdown significantly reduced J399EM (genotype 2a) HCV propagation at the assembly step but not the entry, RNA replication, and protein translation steps of the life cycle. Protein localization and interaction analysis further revealed a role of PLA1A in the interaction of NS2-E2 and NS2-NS5A, as the formation of the NS2-E2 and NS2-NS5A complexes was weakened in the absence of PLA1A. In addition, PLA1A stabilized the NS2/NS5A dotted structure during infection. These data suggest that PLA1A plays an important role in bridging the membrane-associated NS2-E2 complex and the NS5A-associated replication complex via its interaction with E2, NS2, and NS5A, which leads to a coordinating interaction between the structural and nonstructural proteins and facilitates viral assembly.
IMPORTANCEHepatitis C virus (HCV) genomic replication is driven by the replication complex and occurs at the membranous web, while the lipid droplet is the organelle in which virion assembly is initiated. In this study, we identified phosphatidylserine-specific phospholipase A1 (PLA1A), a member of phospholipase A 1 family, as a novel host factor involved in the assembly process of HCV. PLA1A, which is induced by HCV infection at a late infection stage, interacts with HCV E2, NS2, and NS5A proteins and enhances and stabilizes the NS2-E2 and NS2-NS5A complex formation, which is essential for viral assembly. Thus, PLA1A is an important host factor which is involved in the initiation of the viral assembly in close proximity to Core-decorated lipid droplets through bringing together the HCV replication complex and envelope complex.
Hepatitis C virus (HCV) is a major cause of chronic liver disease, affecting approximately 185 million people worldwide (1). HCV is a positive single-stranded RNA virus belonging to the Flaviviridae family. The HCV 9.6-kb genome contains a large open reading frame encoding a single polyprotein that is processed into its structural proteins (Core, E1, and E2) and nonstructural (NS) proteins (p7, NS2, NS3, NS4A, NS4B, NS5A, and NS5B) by host and viral proteinases (2). The structural proteins are components of the virion, while nonstructural proteins NS3 to NS5B compose the minimal viral replicase governing RNA replication (3, 4).The overall HCV life cycle has been well defined since the development of an infectious HCV cell culture system (5-7). HCV genomic replication is driven by the replication complex (RC) and occurs at the membranous web, a rearranged membrane structure induced by virus infection (8-10). Recent progress regarding the study of the assembly process demonstrates that the lipid droplet (LD) is the organelle in which virion assembly is initiated (11) and that, in addition to the structural prote...