Complex Processing and Protein:Protein Interactions in the E2:NS2 Region of HCV

Hepatitis C virus (HCV), the principal cause of parenteral non-A, non-B hepatitis, is an RNA virus and a member of the Flaviviridae family. Its genome is translated into a single polyprotein that is processed co- and post-translationally into both structural and nonstructural (NS) proteins There are three putative structural proteins, consisting of the nucleocapsid protein and two envelope glycoproteins, E1 and E2. Analysis of transient transfections of serially extended templates covering the E2/NS2 region provided evidence for three E2 species with distinct C-termini. One form is E2 terminating at amino acid 729, while the larger two species represent fusions with the downstream NS2A and NS2A/NS2B proteins terminating at amino acids 809 and 1026, respectively. Using the same E2 templates, we defined a region of E2 important for co-immunoprecipitation of E1 and observed that this region also prevents E2 secretion. The N-terminus of NS2B was determined by radiosequencing and a novel association of NS2B and probable NS4B with E2 was observed; the regions of NS2B and E2 important for this association have been mapped. These data indicate that complex processing and protein:protein interactions occult during HCV morphogenesis.