Cell surface expression of herpes simplex virus type 1 glycoprotein H in recombinant baculovirus-infected cells

Virology. 1991 Nov;185(1):187-94. doi: 10.1016/0042-6822(91)90766-5.

Abstract

DNA encoding the complete sequence for the herpes simplex virus type 1 (HSV-1) glycoprotein H (gH) was inserted into a baculovirus transfer vector under control of the baculovirus polyhedrin gene promoter. After co-transfection with wild-type baculovirus DNA, recombinants expressing gH were isolated by plaque purification. The baculovirus-expressed HSV-1 gH represented a significant portion of total cellular protein and was several hundred fold more abundant than gH in HSV-1-infected Vero cells. The expressed gH appeared to be glycosylated, since it was similar in size to wild-type HSV-1 gH, was susceptible to both tunicamycin and endoglycosidase-H treatment, and was labeled by [3H]mannose. In contrast to previous reports of gH expressed in mammalian cells, the baculovirus recombinant-expressed gH was abundant on the cell surface as judged by indirect immunofluorescence. To our knowledge, this is the first report of expressed HSV-1 gH being transported to the cell surface in the absence of other HSV-1 gene products and the first report of expressed gH with an apparent molecular weight similar to authentic HSV-1 gH.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Baculoviridae / genetics*
  • Base Sequence
  • Cell Line
  • Cell Membrane / ultrastructure
  • DNA, Viral / genetics*
  • DNA, Viral / isolation & purification
  • Fluorescent Antibody Technique
  • Gene Expression / drug effects
  • Genetic Vectors
  • Insecta
  • Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase / pharmacology
  • Molecular Sequence Data
  • Recombinant Proteins / analysis
  • Recombinant Proteins / biosynthesis
  • Restriction Mapping
  • Simplexvirus / genetics*
  • Transfection*
  • Tunicamycin / pharmacology
  • Viral Envelope Proteins / analysis
  • Viral Envelope Proteins / biosynthesis
  • Viral Envelope Proteins / genetics*

Substances

  • DNA, Viral
  • Recombinant Proteins
  • Viral Envelope Proteins
  • glycoprotein H, herpes simplex virus type 1
  • Tunicamycin
  • Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase