Coexpression of the membrane glycoproteins G1 and G2 of Hantaan virus is required for targeting to the Golgi complex

Abstract

To study the intracellular transport and targeting to the Golgi complex of the membrane glycoproteins G1 and G2 of Hantaan virus, we have expressed them together and separately using recombinant vaccinia viruses. When expressed from the same recombinant vaccinia virus, G1 and G2 were localized to the Golgi complex as analyzed by both immunofluorescence and subcellular fractionation. However, when the glycoproteins were expressed from separate recombinant viruses, both proteins remained in the endoplasmic reticulum. Using several monoclonal antibodies, it was found that G1 expressed alone did not acquire its correct conformation. Finally, if cells were coinfected with G1- and G2-expressing recombinant viruses, the proteins were again targeted to the Golgi complex. The N-linked glycans remained in all cases largely endoglycosidase-H sensitive. With none of the recombinant viruses were expression of the glycoproteins observed on the cell surface. Neither did chasing in the presence of cycloheximide result in the surface expression of G1 or G2. Our results indicate that for transport out of the endoplasmic reticulum and proper targeting to the Golgi complex, the two glycoproteins have to be coexpressed. The most likely interpretation is that G1 and G2 have to interact with each other in the endoplasmic reticulum in order to become transport competent.