Host-derived apolipoproteins play comparable roles with viral secretory proteins Erns and NS1 in the infectious particle formation of Flaviviridae

Amphipathic α-helices of exchangeable apolipoproteins have shown to play crucial roles in the formation of infectious hepatitis C virus (HCV) particles through the interaction with viral particles. Among the Flaviviridae members, pestivirus and flavivirus possess a viral structural protein Erns or a non-structural protein 1 (NS1) as secretory glycoproteins, respectively, while Hepacivirus including HCV has no secretory glycoprotein. In case of pestivirus replication, the C-terminal long amphipathic α-helices of Erns are important for anchoring to viral membrane. Here we show that host-derived apolipoproteins play functional roles similar to those of virally encoded Erns and NS1 in the formation of infectious particles. We examined whether Erns and NS1 could compensate for the role of apolipoproteins in particle formation of HCV in apolipoprotein B (ApoB) and ApoE double-knockout Huh7 (BE-KO), and non-hepatic 293T cells. We found that exogenous expression of either Erns or NS1 rescued infectious particle formation of HCV in the BE-KO and 293T cells. In addition, expression of apolipoproteins or NS1 partially rescued the production of infectious pestivirus particles in cells upon electroporation with an Erns-deleted non-infectious RNA. As with exchangeable apolipoproteins, the C-terminal amphipathic α-helices of Erns play the functional roles in the formation of infectious HCV or pestivirus particles. These results strongly suggest that the host- and virus-derived secretory glycoproteins have overlapping roles in the viral life cycle of Flaviviridae, especially in the maturation of infectious particles, while Erns and NS1 also participate in replication complex formation and viral entry, respectively. Considering the abundant hepatic expression and liver-specific propagation of these apolipoproteins, HCV might have evolved to utilize them in the formation of infectious particles through deletion of a secretory viral glycoprotein gene.
Author summary The family Flaviviridae consists of 4 genera, namely Flavivirus, Pestivirus, Pegivirus, and Hepacivirus. Flaviviruses and pestiviruses can infect various species and tissues; however, infection of pegivirus and hepacivirus is observed in a strikingly restricted range of tissue and hosts. Although all the Flaviviridae viruses possess a similar genome structure, hepatitis C virus (HCV) from Hepacivirus encodes no secretory glycoprotein, such as Erns of pestivirus and NS1 of flavivirus. The apolipoproteins, one of the host secretory glycoproteins, play important roles in the formation of infectious HCV particles through the interaction with viral particles. The data presented here show that the host-derived apolipoproteins and viral-derived Erns and NS1 have overlapping roles in the maturation of infectious particles of Flaviviridae. Considering an abundant expression of apolipoproteins in the liver and their liver-specific propagation, HCV might have evolved to utilize the apolipoproteins in the formation of infectious particles through deletion of a gene encoding a secretory viral glycoprotein. The data of this manuscript also suggest that utilization of host factors in the viral life cycle is closely associated with the tissue- and species-specificities and evolution among Flaviviridae viruses.