Glycoprotein B (gB) is conserved among the herpesvirus family which infects a broad range of species. To investigate the functional homology of human alpha-herpesviruses, beta-herpesviruses, and gamma-herpesviruses gB proteins, complementation studies were performed with gB genes from each subfamily member using EBV gp110 (EBV gB homologue) and HSV-1 gB null mutants. Neither the alpha-herpesvirus HSV-1 gB gene nor the beta-herpesvirus HCMV gB gene were able to complement the gp110 null mutant. Conversely, neither the beta-herpesvirus HCMV gB or the gamma-herpesvirus EBV gp110 gene were able to complement HSV-1 gB null mutants. To further investigate functional domains of EBV gp110 and HSV-1 gB, gB-gp110 chimeric proteins were constructed. Surprisingly, none of the chimeric proteins were able to complement either HSV-1 gB null mutants or EBV gp110 null mutants. These results demonstrate that there is not sufficient functional homology between the different gBs to allow complementation in other subfamily members of the herpesvirus family. (C) 1997 Academic Press.