[en] Genes encoding glycoprotein gH and gL homologues were localized in the genome of the gamma-herpesvirus bovine herpesvirus-4 (BHV-4). Both genes were sequenced and glutathione S-transferase fusion proteins were produced and used to immunize rabbits against the translation products of the two genes. The anti-gH serum recognized a protein with an apparent molecular mass (MM) of 110 kDa both in infected cells and in virions. This protein was sensitive to endo-beta-N-acetylglucosaminase-H (endoH) and endoglycosidase F-N-glycosidase F (endoF-PNGaseF) digestion. A protein with the same relative mobility was immunoprecipitated from infected cells radiolabelled with [3H]glucosamine which confirmed that this product (gp110), now designated BHV-4 gH, was glycosylated. Western blotting with the anti-gL serum detected in infected cells a product with an apparent MM ranging from 31-35 kDa and diffusely migrating protein species ranging from 45-65 kDa. Tunicamycin, monensin, endoH or endoF-PNGaseF treatments showed that both the 31-35 kDa and the 45-65 kDa proteins were glycosylated, gp31-35 being a precursor of the 45-65 kDa glycoprotein species. In radioimmunoprecipitation assays, the anti-gL serum immunoprecipitated from infected cells two glycosylated proteins with apparent MMs of 31-35 kDa (gp31-35) and 45-55 kDa (gp45-55). However a third glycoprotein, gp110, was also immunoprecipitated together with gp31-35 and gp45-55. gp110 and gp45-55 were subsequently confirmed to be virion glycoproteins corresponding to mature forms of BHV-4 gH and gL respectively. In addition, the present study clearly demonstrated complex formation between BHV-4 gH and gL both in virions and in infected cells.
Disciplines :
Microbiology Veterinary medicine & animal health
Author, co-author :
Lomonte, P.
Filée, Patrice ; Université de Liège - ULiège > Centre d'ingénierie des protéines
Lyaku, J. R.
Bublot, M.
Pastoret, Paul-Pierre ; Université de Liège - ULiège > Département des maladies infectieuses et parasitaires > Département des maladies infectieuses et parasitaires
Thiry, Etienne ; Université de Liège - ULiège > Département des maladies infectieuses et parasitaires > Virologie, épidémiologie et pathologie des maladies virales
Language :
English
Title :
Analysis of the Biochemical Properties of, and Complex Formation between, Glycoproteins H and L of the Gamma2 Herpesvirus Bovine Herpesvirus-4
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