Characterization of herpes simplex virus type 1 recombinants that express and incorporate high levels of HCV E2-gC chimeric proteins.
We report the construction of two HSV-1 recombinants encoding chimeric forms of the E2 glycoprotein of HCV-1a composed of the ectodomain of E2 (aa384-611 or 384-711) fused to different parts of the transmembrane and cytoplasmic domain of the HSV-1 gC glycoprotein (gC). The parental HSV-1, known as KgBpK(-)gC(-), is deleted for gC and the main heparan sulphate (HS) binding domain of gB, and it exhibits impaired binding (ca. 80%) to HS compared to the wild type virus KOS [Laquerre, S., Argnani, R., Anderson, D.B., Zucchini, S., Manservigi, R., Glorioso, J.C., 1998. Heparan sulphate proteoglycan binding by herpes simplex virus type 1 glycoproteins B and C, which differ in their contributions to virus attachment, penetration, and cell-to-cell spread. J. Virol. 72, 6119-6130]. We show that gC:E2 proteins are efficiently expressed and transported to the cell surface. We also demonstrate that HSV-1 can incorporate both gC:E2 chimeric proteins into particles and show that incorporation of both chimeric molecules in the viral envelope partially restored binding (ca. 20%) of the HSV-1 recombinants to heparan sulphate. Finally, we showed that the gC:E2ScaI chimeric glycoprotein was able to bind a recombinant form of hCD81 and virion-expressed gC:E2ScaI permitted the binding of the HSV-1 recombinant virus to the hCD81 molecule.
Kouvatsis
V
V
Molecular Virology Laboratory, Hellenic Pasteur Institute, Athens 11521, Greece.
Argnani
R
R
Tsitoura
E
E
Arsenakis
M
M
Georgopoulou
U
U
Mavromara
P
P
Manservigi
R
R
D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D002460:Cell Line; D002522:Chlorocebus aethiops; D018259:Herpesvirus 1, Human / Q000502:physiology*; D006801:Humans; D017434:Protein Structure, Tertiary; D016865:Reassortant Viruses / Q000502:physiology*; D011991:Receptors, Virus / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis; D060225:Tetraspanin 28; D014709:Vero Cells; D014759:Viral Envelope Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D014779:Virus Replication
D015703:Antigens, CD; C554798:CD81 protein, human; D011991:Receptors, Virus; D011994:Recombinant Proteins; D060225:Tetraspanin 28; D014759:Viral Envelope Proteins; C038535:glycoprotein gC, herpes simplex virus type 1; C091015:glycoprotein E2, Hepatitis C virus