Four consecutive arginine residues at positions 836-839 of EBV gp110 determine intracellular localization of gp110.

Epstein-Barr virus (EBV) glycoprotein 110 (gp110) has sequence homology with herpes simplex virus-1 (HSV-1) gB; however the role of gp110 in EBVs' life cycle differs from that of gB. Unlike HSV-1 gB, which is essential for HSV-1 infection but dispensable for virus production, gp110 is required for assembly and egress of EBV. EBV gp110 is found mainly in the endoplasmic reticulum (ER)/nuclear membrane, whereas little or no gp110 is detected in the plasma membrane or a mature viral particle. Conversely, HSV-1 gB is abundant in the envelope of mature virions and in the plasma membrane as well as in the ER/nuclear membrane of HSV-1-infected cells. Interestingly, there are four consecutive arginine residues (at positions 836-839 of gp110) in the C-terminal domain previously shown to be important for gp110's intracellular localization. To determine whether these arginines function as an ER/nuclear localization signal, point mutants were constructed differentially substituting the four arginines. The glycosylation pattern and intracellular localization of the mutants were investigated by assessing sensitivity to endoglycosidase H (endo H) digestion and performing indirect immunofluorescence assays. Substitution of part of the four arginines changed the glycosylation profile and targeting of gp110. In addition, mutations preserving the net charge of the four arginines as well as those causing net charge shift resulted in the changed intracellular localization and altered glycosylation pattern. These results suggest that not only the net charge but also the conformation of the four arginines are important for gp110's processing and subcellular localization. Copyright 1999 Academic Press.