Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1995 Nov;69(11):7362–7366. doi: 10.1128/jvi.69.11.7362-7366.1995

Herpes simplex virus capsids assembled in insect cells infected with recombinant baculoviruses: structural authenticity and localization of VP26.

B L Trus 1, F L Homa 1, F P Booy 1, W W Newcomb 1, D R Thomsen 1, N Cheng 1, J C Brown 1, A C Steven 1
PMCID: PMC189670  PMID: 7474170

Abstract

Recently, recombinant baculoviruses have been used to show that expression of six herpes simplex virus type 1 genes results in the formation of capsid-like particles. We have applied cryoelectron microscopy and three-dimensional image reconstruction to establish their structural authenticity to a resolution of approximately 2.7 nm. By comparing capsids assembled with and without the expression of gene UL35, we have confirmed the presence of six copies of its product, VP26 (12 kDa), around each hexon tip. However, VP26 is not present on pentons, indicating that the conformational differences between the hexon and penton states of the major capsid protein, VP5, extend to the VP26 binding site.

Full Text

The Full Text of this article is available as a PDF (2.7 MB).

Selected References

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

  1. Baker T. S., Drak J., Bina M. The capsid of small papova viruses contains 72 pentameric capsomeres: direct evidence from cryo-electron-microscopy of simian virus 40. Biophys J. 1989 Feb;55(2):243–253. doi: 10.1016/S0006-3495(89)82799-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baker T. S., Newcomb W. W., Booy F. P., Brown J. C., Steven A. C. Three-dimensional structures of maturable and abortive capsids of equine herpesvirus 1 from cryoelectron microscopy. J Virol. 1990 Feb;64(2):563–573. doi: 10.1128/jvi.64.2.563-573.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Booy F. P., Newcomb W. W., Trus B. L., Brown J. C., Baker T. S., Steven A. C. Liquid-crystalline, phage-like packing of encapsidated DNA in herpes simplex virus. Cell. 1991 Mar 8;64(5):1007–1015. doi: 10.1016/0092-8674(91)90324-r. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Booy F. P., Trus B. L., Newcomb W. W., Brown J. C., Conway J. F., Steven A. C. Finding a needle in a haystack: detection of a small protein (the 12-kDa VP26) in a large complex (the 200-MDa capsid of herpes simplex virus). Proc Natl Acad Sci U S A. 1994 Jun 7;91(12):5652–5656. doi: 10.1073/pnas.91.12.5652. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Casjens S., King J. Virus assembly. Annu Rev Biochem. 1975;44:555–611. doi: 10.1146/annurev.bi.44.070175.003011. [DOI] [PubMed] [Google Scholar]
  6. Cheng R. H., Reddy V. S., Olson N. H., Fisher A. J., Baker T. S., Johnson J. E. Functional implications of quasi-equivalence in a T = 3 icosahedral animal virus established by cryo-electron microscopy and X-ray crystallography. Structure. 1994 Apr 15;2(4):271–282. doi: 10.1016/s0969-2126(00)00029-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Conway J. F., Trus B. L., Booy F. P., Newcomb W. W., Brown J. C., Steven A. C. The effects of radiation damage on the structure of frozen hydrated HSV-1 capsids. J Struct Biol. 1993 Nov-Dec;111(3):222–233. doi: 10.1006/jsbi.1993.1052. [DOI] [PubMed] [Google Scholar]
  8. Crowther R. A. Procedures for three-dimensional reconstruction of spherical viruses by Fourier synthesis from electron micrographs. Philos Trans R Soc Lond B Biol Sci. 1971 May 27;261(837):221–230. doi: 10.1098/rstb.1971.0054. [DOI] [PubMed] [Google Scholar]
  9. Davison M. D., Rixon F. J., Davison A. J. Identification of genes encoding two capsid proteins (VP24 and VP26) of herpes simplex virus type 1. J Gen Virol. 1992 Oct;73(Pt 10):2709–2713. doi: 10.1099/0022-1317-73-10-2709. [DOI] [PubMed] [Google Scholar]
  10. Friedmann A., Coward J. E., Rosenkranz H. S., Morgan C. Electron microscopic studies on assembly of herpes simplex virus upon removal of hydroxyurea block. J Gen Virol. 1975 Feb;26(2):171–181. doi: 10.1099/0022-1317-26-2-171. [DOI] [PubMed] [Google Scholar]
  11. Furlong D. Direct evidence for 6-fold symmetry of the herpesvirus hexon capsomere. Proc Natl Acad Sci U S A. 1978 Jun;75(6):2764–2766. doi: 10.1073/pnas.75.6.2764. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. McNabb D. S., Courtney R. J. Identification and characterization of the herpes simplex virus type 1 virion protein encoded by the UL35 open reading frame. J Virol. 1992 May;66(5):2653–2663. doi: 10.1128/jvi.66.5.2653-2663.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Newcomb W. W., Brown J. C., Booy F. P., Steven A. C. Nucleocapsid mass and capsomer protein stoichiometry in equine herpesvirus 1: scanning transmission electron microscopic study. J Virol. 1989 Sep;63(9):3777–3783. doi: 10.1128/jvi.63.9.3777-3783.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Newcomb W. W., Homa F. L., Thomsen D. R., Ye Z., Brown J. C. Cell-free assembly of the herpes simplex virus capsid. J Virol. 1994 Sep;68(9):6059–6063. doi: 10.1128/jvi.68.9.6059-6063.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Newcomb W. W., Trus B. L., Booy F. P., Steven A. C., Wall J. S., Brown J. C. Structure of the herpes simplex virus capsid. Molecular composition of the pentons and the triplexes. J Mol Biol. 1993 Jul 20;232(2):499–511. doi: 10.1006/jmbi.1993.1406. [DOI] [PubMed] [Google Scholar]
  16. Palmer E. L., Martin M. L., Gary G. W., Jr The ultrastructure of disrupted herpesvirus nucleocapsids. Virology. 1975 May;65(1):260–265. doi: 10.1016/0042-6822(75)90026-4. [DOI] [PubMed] [Google Scholar]
  17. Patel A. H., MacLean J. B. The product of the UL6 gene of herpes simplex virus type 1 is associated with virus capsids. Virology. 1995 Jan 10;206(1):465–478. doi: 10.1016/s0042-6822(95)80062-x. [DOI] [PubMed] [Google Scholar]
  18. Schrag J. D., Prasad B. V., Rixon F. J., Chiu W. Three-dimensional structure of the HSV1 nucleocapsid. Cell. 1989 Feb 24;56(4):651–660. doi: 10.1016/0092-8674(89)90587-4. [DOI] [PubMed] [Google Scholar]
  19. Steven A. C., Roberts C. R., Hay J., Bisher M. E., Pun T., Trus B. L. Hexavalent capsomers of herpes simplex virus type 2: symmetry, shape, dimensions, and oligomeric status. J Virol. 1986 Feb;57(2):578–584. doi: 10.1128/jvi.57.2.578-584.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Tatman J. D., Preston V. G., Nicholson P., Elliott R. M., Rixon F. J. Assembly of herpes simplex virus type 1 capsids using a panel of recombinant baculoviruses. J Gen Virol. 1994 May;75(Pt 5):1101–1113. doi: 10.1099/0022-1317-75-5-1101. [DOI] [PubMed] [Google Scholar]
  21. Thomsen D. R., Newcomb W. W., Brown J. C., Homa F. L. Assembly of the herpes simplex virus capsid: requirement for the carboxyl-terminal twenty-five amino acids of the proteins encoded by the UL26 and UL26.5 genes. J Virol. 1995 Jun;69(6):3690–3703. doi: 10.1128/jvi.69.6.3690-3703.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Thomsen D. R., Roof L. L., Homa F. L. Assembly of herpes simplex virus (HSV) intermediate capsids in insect cells infected with recombinant baculoviruses expressing HSV capsid proteins. J Virol. 1994 Apr;68(4):2442–2457. doi: 10.1128/jvi.68.4.2442-2457.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Trus B. L., Newcomb W. W., Booy F. P., Brown J. C., Steven A. C. Distinct monoclonal antibodies separately label the hexons or the pentons of herpes simplex virus capsid. Proc Natl Acad Sci U S A. 1992 Dec 1;89(23):11508–11512. doi: 10.1073/pnas.89.23.11508. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. WILDY P., RUSSELL W. C., HORNE R. W. The morphology of herpes virus. Virology. 1960 Oct;12:204–222. doi: 10.1016/0042-6822(60)90195-1. [DOI] [PubMed] [Google Scholar]
  25. Zhou Z. H., Prasad B. V., Jakana J., Rixon F. J., Chiu W. Protein subunit structures in the herpes simplex virus A-capsid determined from 400 kV spot-scan electron cryomicroscopy. J Mol Biol. 1994 Sep 30;242(4):456–469. doi: 10.1006/jmbi.1994.1594. [DOI] [PubMed] [Google Scholar]

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

RESOURCES