Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1991 Aug;65(8):4275–4283. doi: 10.1128/jvi.65.8.4275-4283.1991

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

S L Highlander 1, W F Goins 1, S Person 1, T C Holland 1, M Levine 1, J C Glorioso 1
PMCID: PMC248865  PMID: 1649330

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 cell 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.

Full text

PDF

Images in this article

4277Image
on p.4277
4277Image
on p.4277
4278Image
on p.4278
4278Image
on p.4278
4279Image
on p.4279
4279Image
on p.4279

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ali M. A. Oligomerization of herpes simplex virus glycoprotein B occurs in the endoplasmic reticulum and a 102 amino acid cytosolic domain is dispensable for dimer assembly. Virology. 1990 Oct;178(2):588–592. doi: 10.1016/0042-6822(90)90359-y. [DOI] [PubMed] [Google Scholar]
  2. Buckmaster E. A., Gompels U., Minson A. Characterisation and physical mapping of an HSV-1 glycoprotein of approximately 115 X 10(3) molecular weight. Virology. 1984 Dec;139(2):408–413. doi: 10.1016/0042-6822(84)90387-8. [DOI] [PubMed] [Google Scholar]
  3. Bzik D. J., Debroy C., Fox B. A., Pederson N. E., Person S. The nucleotide sequence of the gB glycoprotein gene of HSV-2 and comparison with the corresponding gene of HSV-1. Virology. 1986 Dec;155(2):322–333. doi: 10.1016/0042-6822(86)90196-0. [DOI] [PubMed] [Google Scholar]
  4. Bzik D. J., Fox B. A., DeLuca N. A., Person S. Nucleotide sequence of a region of the herpes simplex virus type 1 gB glycoprotein gene: mutations affecting rate of virus entry and cell fusion. Virology. 1984 Aug;137(1):185–190. doi: 10.1016/0042-6822(84)90022-9. [DOI] [PubMed] [Google Scholar]
  5. Bzik D. J., Fox B. A., DeLuca N. A., Person S. Nucleotide sequence specifying the glycoprotein gene, gB, of herpes simplex virus type 1. Virology. 1984 Mar;133(2):301–314. doi: 10.1016/0042-6822(84)90397-0. [DOI] [PubMed] [Google Scholar]
  6. Cai W. H., Gu B., Person S. Role of glycoprotein B of herpes simplex virus type 1 in viral entry and cell fusion. J Virol. 1988 Aug;62(8):2596–2604. doi: 10.1128/jvi.62.8.2596-2604.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cai W. Z., Person S., DebRoy C., Gu B. H. Functional regions and structural features of the gB glycoprotein of herpes simplex virus type 1. An analysis of linker insertion mutants. J Mol Biol. 1988 Jun 5;201(3):575–588. doi: 10.1016/0022-2836(88)90639-0. [DOI] [PubMed] [Google Scholar]
  8. Cai W. Z., Person S., Warner S. C., Zhou J. H., DeLuca N. A. Linker-insertion nonsense and restriction-site deletion mutations of the gB glycoprotein gene of herpes simplex virus type 1. J Virol. 1987 Mar;61(3):714–721. doi: 10.1128/jvi.61.3.714-721.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Chakrabarti S., Mizukami T., Franchini G., Moss B. Synthesis, oligomerization, and biological activity of the human immunodeficiency virus type 2 envelope glycoprotein expressed by a recombinant vaccinia virus. Virology. 1990 Sep;178(1):134–142. doi: 10.1016/0042-6822(90)90386-6. [DOI] [PubMed] [Google Scholar]
  10. Chapsal J. M., Pereira L. Characterization of epitopes on native and denatured forms of herpes simplex virus glycoprotein B. Virology. 1988 Jun;164(2):427–434. doi: 10.1016/0042-6822(88)90556-9. [DOI] [PubMed] [Google Scholar]
  11. Claesson-Welsh L., Spear P. G. Amino-terminal sequence, synthesis, and membrane insertion of glycoprotein B of herpes simplex virus type 1. J Virol. 1987 Jan;61(1):1–7. doi: 10.1128/jvi.61.1.1-7.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Claesson-Welsh L., Spear P. G. Oligomerization of herpes simplex virus glycoprotein B. J Virol. 1986 Nov;60(2):803–806. doi: 10.1128/jvi.60.2.803-806.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Copeland C. S., Zimmer K. P., Wagner K. R., Healey G. A., Mellman I., Helenius A. Folding, trimerization, and transport are sequential events in the biogenesis of influenza virus hemagglutinin. Cell. 1988 Apr 22;53(2):197–209. doi: 10.1016/0092-8674(88)90381-9. [DOI] [PubMed] [Google Scholar]
  14. Cranage M. P., Kouzarides T., Bankier A. T., Satchwell S., Weston K., Tomlinson P., Barrell B., Hart H., Bell S. E., Minson A. C. Identification of the human cytomegalovirus glycoprotein B gene and induction of neutralizing antibodies via its expression in recombinant vaccinia virus. EMBO J. 1986 Nov;5(11):3057–3063. doi: 10.1002/j.1460-2075.1986.tb04606.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. DeLuca N., Bzik D. J., Bond V. C., Person S., Snipes W. Nucleotide sequences of herpes simplex virus type 1 (HSV-1) affecting virus entry, cell fusion, and production of glycoprotein gb (VP7). Virology. 1982 Oct 30;122(2):411–423. doi: 10.1016/0042-6822(82)90240-9. [DOI] [PubMed] [Google Scholar]
  16. Desai P. J., Schaffer P. A., Minson A. C. Excretion of non-infectious virus particles lacking glycoprotein H by a temperature-sensitive mutant of herpes simplex virus type 1: evidence that gH is essential for virion infectivity. J Gen Virol. 1988 Jun;69(Pt 6):1147–1156. doi: 10.1099/0022-1317-69-6-1147. [DOI] [PubMed] [Google Scholar]
  17. Earl P. L., Doms R. W., Moss B. Oligomeric structure of the human immunodeficiency virus type 1 envelope glycoprotein. Proc Natl Acad Sci U S A. 1990 Jan;87(2):648–652. doi: 10.1073/pnas.87.2.648. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Eberle R., Black D., Hilliard J. K. Relatedness of glycoproteins expressed on the surface of simian herpes-virus virions and infected cells to specific HSV glycoproteins. Arch Virol. 1989;109(3-4):233–252. doi: 10.1007/BF01311084. [DOI] [PubMed] [Google Scholar]
  19. Einfeld D., Hunter E. Oligomeric structure of a prototype retrovirus glycoprotein. Proc Natl Acad Sci U S A. 1988 Nov;85(22):8688–8692. doi: 10.1073/pnas.85.22.8688. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Feenstra V., Hodaie M., Johnson D. C. Deletions in herpes simplex virus glycoprotein D define nonessential and essential domains. J Virol. 1990 May;64(5):2096–2102. doi: 10.1128/jvi.64.5.2096-2102.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Gething M. J., McCammon K., Sambrook J. Expression of wild-type and mutant forms of influenza hemagglutinin: the role of folding in intracellular transport. Cell. 1986 Sep 12;46(6):939–950. doi: 10.1016/0092-8674(86)90076-0. [DOI] [PubMed] [Google Scholar]
  22. Gompels U., Minson A. The properties and sequence of glycoprotein H of herpes simplex virus type 1. Virology. 1986 Sep;153(2):230–247. doi: 10.1016/0042-6822(86)90026-7. [DOI] [PubMed] [Google Scholar]
  23. Highlander S. L., Cai W. H., Person S., Levine M., Glorioso J. C. Monoclonal antibodies define a domain on herpes simplex virus glycoprotein B involved in virus penetration. J Virol. 1988 Jun;62(6):1881–1888. doi: 10.1128/jvi.62.6.1881-1888.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Highlander S. L., Dorney D. J., Gage P. J., Holland T. C., Cai W., Person S., Levine M., Glorioso J. C. Identification of mar mutations in herpes simplex virus type 1 glycoprotein B which alter antigenic structure and function in virus penetration. J Virol. 1989 Feb;63(2):730–738. doi: 10.1128/jvi.63.2.730-738.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Highlander S. L., Sutherland S. L., Gage P. J., Johnson D. C., Levine M., Glorioso J. C. Neutralizing monoclonal antibodies specific for herpes simplex virus glycoprotein D inhibit virus penetration. J Virol. 1987 Nov;61(11):3356–3364. doi: 10.1128/jvi.61.11.3356-3364.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Holland T. C., Marlin S. D., Levine M., Glorioso J. Antigenic variants of herpes simplex virus selected with glycoprotein-specific monoclonal antibodies. J Virol. 1983 Feb;45(2):672–682. doi: 10.1128/jvi.45.2.672-682.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Keller P. M., Davison A. J., Lowe R. S., Riemen M. W., Ellis R. W. Identification and sequence of the gene encoding gpIII, a major glycoprotein of varicella-zoster virus. Virology. 1987 Apr;157(2):526–533. doi: 10.1016/0042-6822(87)90295-9. [DOI] [PubMed] [Google Scholar]
  28. Kreis T. E., Lodish H. F. Oligomerization is essential for transport of vesicular stomatitis viral glycoprotein to the cell surface. Cell. 1986 Sep 12;46(6):929–937. doi: 10.1016/0092-8674(86)90075-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Ligas M. W., Johnson D. C. A herpes simplex virus mutant in which glycoprotein D sequences are replaced by beta-galactosidase sequences binds to but is unable to penetrate into cells. J Virol. 1988 May;62(5):1486–1494. doi: 10.1128/jvi.62.5.1486-1494.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Longnecker R., Chatterjee S., Whitley R. J., Roizman B. Identification of a herpes simplex virus 1 glycoprotein gene within a gene cluster dispensable for growth in cell culture. Proc Natl Acad Sci U S A. 1987 Jun;84(12):4303–4307. doi: 10.1073/pnas.84.12.4303. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Longnecker R., Roizman B. Clustering of genes dispensable for growth in culture in the S component of the HSV-1 genome. Science. 1987 May 1;236(4801):573–576. doi: 10.1126/science.3033823. [DOI] [PubMed] [Google Scholar]
  32. Manservigi R., Spear P. G., Buchan A. Cell fusion induced by herpes simplex virus is promoted and suppressed by different viral glycoproteins. Proc Natl Acad Sci U S A. 1977 Sep;74(9):3913–3917. doi: 10.1073/pnas.74.9.3913. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Marlin S. D., Holland T. C., Levine M., Glorioso J. C. Epitopes of herpes simplex virus type 1 glycoprotein gC are clustered in two distinct antigenic sites. J Virol. 1985 Jan;53(1):128–136. doi: 10.1128/jvi.53.1.128-136.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. McGeoch D. J., Dalrymple M. A., Davison A. J., Dolan A., Frame M. C., McNab D., Perry L. J., Scott J. E., Taylor P. The complete DNA sequence of the long unique region in the genome of herpes simplex virus type 1. J Gen Virol. 1988 Jul;69(Pt 7):1531–1574. doi: 10.1099/0022-1317-69-7-1531. [DOI] [PubMed] [Google Scholar]
  35. McGeoch D. J., Dolan A., Donald S., Rixon F. J. Sequence determination and genetic content of the short unique region in the genome of herpes simplex virus type 1. J Mol Biol. 1985 Jan 5;181(1):1–13. doi: 10.1016/0022-2836(85)90320-1. [DOI] [PubMed] [Google Scholar]
  36. Mester J. C., Highlander S. L., Osmand A. P., Glorioso J. C., Rouse B. T. Herpes simplex virus type 1-specific immunity induced by peptides corresponding to an antigenic site of glycoprotein B. J Virol. 1990 Nov;64(11):5277–5283. doi: 10.1128/jvi.64.11.5277-5283.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Misra V., Nelson R., Smith M. Sequence of a bovine herpesvirus type-1 glycoprotein gene that is homologous to the herpes simplex gene for the glycoprotein gB. Virology. 1988 Oct;166(2):542–549. doi: 10.1016/0042-6822(88)90525-9. [DOI] [PubMed] [Google Scholar]
  38. Noble A. G., Lee G. T., Sprague R., Parish M. L., Spear P. G. Anti-gD monoclonal antibodies inhibit cell fusion induced by herpes simplex virus type 1. Virology. 1983 Aug;129(1):218–224. doi: 10.1016/0042-6822(83)90409-9. [DOI] [PubMed] [Google Scholar]
  39. Parks G. D., Lamb R. A. Folding and oligomerization properties of a soluble and secreted form of the paramyxovirus hemagglutinin-neuraminidase glycoprotein. Virology. 1990 Oct;178(2):498–508. doi: 10.1016/0042-6822(90)90347-t. [DOI] [PubMed] [Google Scholar]
  40. Pellett P. E., Biggin M. D., Barrell B., Roizman B. Epstein-Barr virus genome may encode a protein showing significant amino acid and predicted secondary structure homology with glycoprotein B of herpes simplex virus 1. J Virol. 1985 Dec;56(3):807–813. doi: 10.1128/jvi.56.3.807-813.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Pellett P. E., Kousoulas K. G., Pereira L., Roizman B. Anatomy of the herpes simplex virus 1 strain F glycoprotein B gene: primary sequence and predicted protein structure of the wild type and of monoclonal antibody-resistant mutants. J Virol. 1985 Jan;53(1):243–253. doi: 10.1128/jvi.53.1.243-253.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Rey M. A., Krust B., Laurent A. G., Montagnier L., Hovanessian A. G. Characterization of human immunodeficiency virus type 2 envelope glycoproteins: dimerization of the glycoprotein precursor during processing. J Virol. 1989 Feb;63(2):647–658. doi: 10.1128/jvi.63.2.647-658.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Richman D. D., Buckmaster A., Bell S., Hodgman C., Minson A. C. Identification of a new glycoprotein of herpes simplex virus type 1 and genetic mapping of the gene that codes for it. J Virol. 1986 Feb;57(2):647–655. doi: 10.1128/jvi.57.2.647-655.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Riggio M. P., Cullinane A. A., Onions D. E. Identification and nucleotide sequence of the glycoprotein gB gene of equine herpesvirus 4. J Virol. 1989 Mar;63(3):1123–1133. doi: 10.1128/jvi.63.3.1123-1133.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Robbins A. K., Dorney D. J., Wathen M. W., Whealy M. E., Gold C., Watson R. J., Holland L. E., Weed S. D., Levine M., Glorioso J. C. The pseudorabies virus gII gene is closely related to the gB glycoprotein gene of herpes simplex virus. J Virol. 1987 Sep;61(9):2691–2701. doi: 10.1128/jvi.61.9.2691-2701.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Ross L. J., Sanderson M., Scott S. D., Binns M. M., Doel T., Milne B. Nucleotide sequence and characterization of the Marek's disease virus homologue of glycoprotein B of herpes simplex virus. J Gen Virol. 1989 Jul;70(Pt 7):1789–1804. doi: 10.1099/0022-1317-70-7-1789. [DOI] [PubMed] [Google Scholar]
  47. Sarmiento M., Spear P. G. Membrane proteins specified by herpes simplex viruses. IV. Conformation of the virion glycoprotein designated VP7(B2). J Virol. 1979 Mar;29(3):1159–1167. doi: 10.1128/jvi.29.3.1159-1167.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Schaffer P. A., Aron G. M., Biswal N., Benyesh-Melnick M. Temperature-sensitive mutants of herpes simplex virus type 1: isolation, complementation and partial characterization. Virology. 1973 Mar;52(1):57–71. doi: 10.1016/0042-6822(73)90398-x. [DOI] [PubMed] [Google Scholar]
  49. Schaffer P. A., Carter V. C., Timbury M. C. Collaborative complementation study of temperature-sensitive mutants of herpes simplex virus types 1 and 2. J Virol. 1978 Sep;27(3):490–504. doi: 10.1128/jvi.27.3.490-504.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Sechoy O., Philippot J. R., Bienvenue A. F protein-F protein interaction within the Sendai virus identified by native bonding or chemical cross-linking. J Biol Chem. 1987 Aug 25;262(24):11519–11523. [PubMed] [Google Scholar]
  51. Stannard L. M., Fuller A. O., Spear P. G. Herpes simplex virus glycoproteins associated with different morphological entities projecting from the virion envelope. J Gen Virol. 1987 Mar;68(Pt 3):715–725. doi: 10.1099/0022-1317-68-3-715. [DOI] [PubMed] [Google Scholar]
  52. Stuve L. L., Brown-Shimer S., Pachl C., Najarian R., Dina D., Burke R. L. Structure and expression of the herpes simplex virus type 2 glycoprotein gB gene. J Virol. 1987 Feb;61(2):326–335. doi: 10.1128/jvi.61.2.326-335.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Weiss C. D., Levy J. A., White J. M. Oligomeric organization of gp120 on infectious human immunodeficiency virus type 1 particles. J Virol. 1990 Nov;64(11):5674–5677. doi: 10.1128/jvi.64.11.5674-5677.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Whalley J. M., Robertson G. R., Scott N. A., Hudson G. C., Bell C. W., Woodworth L. M. Identification and nucleotide sequence of a gene in equine herpesvirus 1 analogous to the herpes simplex virus gene encoding the major envelope glycoprotein gB. J Gen Virol. 1989 Feb;70(Pt 2):383–394. doi: 10.1099/0022-1317-70-2-383. [DOI] [PubMed] [Google Scholar]
  55. Whealy M. E., Robbins A. K., Enquist L. W. The export pathway of the pseudorabies virus gB homolog gII involves oligomer formation in the endoplasmic reticulum and protease processing in the Golgi apparatus. J Virol. 1990 May;64(5):1946–1955. doi: 10.1128/jvi.64.5.1946-1955.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Whitbeck J. C., Bello L. J., Lawrence W. C. Comparison of the bovine herpesvirus 1 gI gene and the herpes simplex virus type 1 gB gene. J Virol. 1988 Sep;62(9):3319–3327. doi: 10.1128/jvi.62.9.3319-3327.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES