The extracellular part of glycoprotein E of bovine herpesvirus 1 is sufficient for complex formation with glycoprotein I but not for cell-to-cell spread
J. Tyborowska et al., The extracellular part of glycoprotein E of bovine herpesvirus 1 is sufficient for complex formation with glycoprotein I but not for cell-to-cell spread, ARCH VIROL, 145(2), 2000, pp. 333-351
Glycoproteins gE and gI of bovine herpesvirus 1 (BHV-1) are type I transmem
brane proteins that can form a complex that is involved in cell-to-cell spr
ead mechanisms. The extracellular domains of both proteins have cysteine-ri
ch regions that are also found in the homologous proteins of other alpha-he
rpesviruses. The extracellular domain of gE has two conserved cysteine-rich
regions: C1 and C2. The other conserved regions in gE are located between
C2 and transmembrane region and in the cytoplasmic domain of gE. We studied
the complex formation between gE and gI using a series of truncated gE pro
teins and a full length form and a secreted form of gI. All proteins were e
xpressed in recombinant baculoviruses. To analyse the compiler formation be
tween these polypeptides we used monoclonal antibodies (MAbs 67 and 75) tha
t specifically react with the gE/gI complex and not with separately express
ed glycoproteins gE and gI alone. This analysis showed that the BHV-1 gE/gI
complex can be formed in insect cells after a co-infection with baculoviru
ses expressing gE and gI in their full length form. When secreted forms of
gE and gI were expressed after co-infection, the gE/gI complex was still fo
rmed and could also be detected in the tissue culture medium. This gE/gI co
mplex was also formed after mixing the tissue culture media of insect cells
expressing the secreted form or gE or gI separately. The smallest part of
gE that still formed a complex is encoded by the first 246 residues of gE.
This extracellular domain contains only the C1 region, showing that the C2
region is not essential for gE/gI complex formation. Shorter forms of gE en
coding the C1 region did not form a detectable complex. We also found that
the formation of gE/gI complex is not sufficient for normal cell-to-cell sp
read of BHV-1. A recombinant BHV-1 gE TM-virus, expressing a truncated glyc
oprotein E from which the transmembrane and cyloplasmic domain were removed
, forms plaques as small as a gE null mutant.