Oligomer formation of the gB glycoprotein of herpes simplex virus type 1

Steven L. Highlander, William F. Goins, Stanley Person, Thomas C. Holland, Myron Levine, Joseph C. Glorioso

Research output: Contribution to journalArticle

Abstract

Oligomer formation of the gB glycoprotein of herpes simplex virus type 1 was studied by sedimentation analysis of radioactively labeled infected cell and virion lysates. Fractions from sucrose gradients were precipitated with a pool of gB-specific monoclonal antibodies and analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Pulse-labeled gB from infected cells was synthesized as monomers and converted to oligomers posttranslationally. The oligomers from infected cells and from virions sedimented as dimers, and there was no evidence of higher-molecular-weight forms. To identify amino acid sequences of gB that contribute to oligomer formation, pairs of mutant plasmids were transfected into Vero cells and superinfected with a gB-null mutant virus to stimulate plasmid-specified gene expression. Radioactively labeled lysates were precipitated with antibodies and examined by SDS-PAGE. Polypeptides from cotransfections were precipitated with an antibody that recognized amino acid sequences present in only one of the two polypeptides. A coprecipitated polypeptide lacking the antibody target epitope was presumed to contain the sequences necessary for oligomer formation. Using this technique, two noncontiguous sites for oligomer formation were detected. An upstream site was localized between residues 93 and 282, and a downstream site was localized between residues 596 and 711. Oligomer formation resulted from molecular interactions between two upstream sites, between two downstream sites, and between an upstream and a downstream site. A schematic diagram of a gB oligomer is presented that is consistent with these data.

Original languageEnglish (US)
Pages (from-to)4275-4283
Number of pages9
JournalJournal of Virology
Volume65
Issue number8
StatePublished - Aug 1991
Externally publishedYes

Fingerprint

Human herpesvirus 1
Human Herpesvirus 1
glycoproteins
Glycoproteins
Sodium Dodecyl Sulfate
Virion
polypeptides
Peptides
Antibodies
Polyacrylamide Gel Electrophoresis
Amino Acid Sequence
Plasmids
virion
antibodies
polyacrylamide gel electrophoresis
plasmids
Vero Cells
amino acid sequences
cells
mutants

ASJC Scopus subject areas

  • Immunology

Cite this

Highlander, S. L., Goins, W. F., Person, S., Holland, T. C., Levine, M., & Glorioso, J. C. (1991). Oligomer formation of the gB glycoprotein of herpes simplex virus type 1. Journal of Virology, 65(8), 4275-4283.

Oligomer formation of the gB glycoprotein of herpes simplex virus type 1. / Highlander, Steven L.; Goins, William F.; Person, Stanley; Holland, Thomas C.; Levine, Myron; Glorioso, Joseph C.

In: Journal of Virology, Vol. 65, No. 8, 08.1991, p. 4275-4283.

Research output: Contribution to journalArticle

Highlander, SL, Goins, WF, Person, S, Holland, TC, Levine, M & Glorioso, JC 1991, 'Oligomer formation of the gB glycoprotein of herpes simplex virus type 1', Journal of Virology, vol. 65, no. 8, pp. 4275-4283.
Highlander SL, Goins WF, Person S, Holland TC, Levine M, Glorioso JC. Oligomer formation of the gB glycoprotein of herpes simplex virus type 1. Journal of Virology. 1991 Aug;65(8):4275-4283.
Highlander, Steven L. ; Goins, William F. ; Person, Stanley ; Holland, Thomas C. ; Levine, Myron ; Glorioso, Joseph C. / Oligomer formation of the gB glycoprotein of herpes simplex virus type 1. In: Journal of Virology. 1991 ; Vol. 65, No. 8. pp. 4275-4283.
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