A single amino acid substitution in the cytoplasmic tail of the glycoprotein B of herpes simplex virus 1 affects both syncytium formation and binding to intracellular heparan sulfate.
Herpes simplex virus 1 (HSV-1) (S) is a spontaneous syncytial mutant derived from the prototype HSV-1(F) after extensive plaque purification, and produces large syncytial plaques on Vero cells. Marker transfer experiments and DNA sequence analysis mapped the syncytial phenotype to a T-C base substitution at codon 787 of the cytoplasmic domain of mature gB, that results in Leu to Pro substitution and consequently belongs to the syn 3 locus. Both the cytoplasmic and the extracellular domains of gB are active in the fusion event since the addition of anti-gB monoclonal antibodies that recognize the extracellular domain of gB prevent HSV-1(S) induced cell fusion. Similarly, gD also participates in cell fusion since addition of anti-gD monoclonal antibodies also prevent HSV-1(S) induced cell fusion. Furthermore the glycoproteins B and D formed complexes in cells infected with mutant or wild type viruses. The amount of gB bound to total heparan sulfate is lower in the mutant than in the wild type strain. This difference becomes particularly profound when gB is associated with a portion of heparan sulfate intercalated to the membranes. The discrepancy in the binding of the mutant and wild type gB to heparan sulfate may be related to the mechanism of cell fusion induced by HSV-1(S).
Diakidi-Kosta
A
A
Laboratory of General Microbiology, Section of Genetics, Development and Molecular Biology, School of Biology, Aristotle University, Thessaloniki 54124, Greece.
Michailidou
G
G
Kontogounis
G
G
Sivropoulou
A
A
Arsenakis
M
M
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D000911:Antibodies, Monoclonal; D001665:Binding Sites; D002522:Chlorocebus aethiops; D002846:Chromatography, Affinity; D003593:Cytoplasm / Q000821:virology; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification; D015726:Giant Cells / Q000502:physiology*; D006497:Heparan Sulfate / Q000378:metabolism*; D018259:Herpesvirus 1, Human / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D017354:Point Mutation; D014162:Transfection; D014709:Vero Cells; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification
D000911:Antibodies, Monoclonal; D004279:DNA, Viral; D006497:Heparan Sulfate; D014759:Viral Envelope Proteins; C040495:glycoprotein B, Simplexvirus