Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1993 Jan;67(1):315–322. doi: 10.1128/jvi.67.1.315-322.1993

Self-assembly of human papillomavirus type 1 capsids by expression of the L1 protein alone or by coexpression of the L1 and L2 capsid proteins.

M E Hagensee 1, N Yaegashi 1, D A Galloway 1
PMCID: PMC237365  PMID: 8380079

Abstract

Vaccinia virus vectors were used to express the major (L1) and minor (L2) capsid proteins of human papillomavirus type 1 (HPV-1) with the vaccinia virus early (p7.5K) or late (pSynth, p11K) promoters. All constructs expressed the appropriate-sized HPV proteins, and both L1 and L2, singly or in combination, localized to the nucleus. Capsids were purified by cesium chloride density gradient centrifugation from nuclei of cells infected with a vaccinia virus-L1 (vac-L1) recombinant or a vac-L1-L2 recombinant but not from vac-L2-infected cells. Electron microscopy showed that the particles were 55 nm in diameter and had icosahedral symmetry. Immunogold-labeled antibodies confirmed the presence of the L1 and L2 proteins in the HPV-1 capsids. Capsids containing L1 alone were fewer and more variable in size and shape than capsids containing the L1 and L2 proteins. The L1-plus-L2 capsids were indistinguishable in appearance from HPV-1 virions obtained from plantar warts. The ability to produce HPV capsids in vitro will be useful in many studies of HPV pathogenicity.

Full text

PDF
315

Images in this article

318Image
on p.318
319Image
on p.319
319Image
on p.319
320Image
on p.320
320Image
on p.320

Selected References

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

  1. Asselineau D., Prunieras M. Reconstruction of 'simplified' skin: control of fabrication. Br J Dermatol. 1984 Jul;111 (Suppl 27):219–222. doi: 10.1111/j.1365-2133.1984.tb15608.x. [DOI] [PubMed] [Google Scholar]
  2. Baker T. S., Drak J., Bina M. Reconstruction of the three-dimensional structure of simian virus 40 and visualization of the chromatin core. Proc Natl Acad Sci U S A. 1988 Jan;85(2):422–426. doi: 10.1073/pnas.85.2.422. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. 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]
  4. Baker T. S., Newcomb W. W., Olson N. H., Cowsert L. M., Olson C., Brown J. C. Structures of bovine and human papillomaviruses. Analysis by cryoelectron microscopy and three-dimensional image reconstruction. Biophys J. 1991 Dec;60(6):1445–1456. doi: 10.1016/S0006-3495(91)82181-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Danos O., Engel L. W., Chen E. Y., Yaniv M., Howley P. M. Comparative analysis of the human type 1a and bovine type 1 papillomavirus genomes. J Virol. 1983 May;46(2):557–566. doi: 10.1128/jvi.46.2.557-566.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Davison A. J., Moss B. New vaccinia virus recombination plasmids incorporating a synthetic late promoter for high level expression of foreign proteins. Nucleic Acids Res. 1990 Jul 25;18(14):4285–4286. doi: 10.1093/nar/18.14.4285. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dollard S. C., Wilson J. L., Demeter L. M., Bonnez W., Reichman R. C., Broker T. R., Chow L. T. Production of human papillomavirus and modulation of the infectious program in epithelial raft cultures. OFF. Genes Dev. 1992 Jul;6(7):1131–1142. doi: 10.1101/gad.6.7.1131. [DOI] [PubMed] [Google Scholar]
  8. Doorbar J., Gallimore P. H. Identification of proteins encoded by the L1 and L2 open reading frames of human papillomavirus 1a. J Virol. 1987 Sep;61(9):2793–2799. doi: 10.1128/jvi.61.9.2793-2799.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Favre M. Structural polypeptides of rabbit, bovine, and human papillomaviruses. J Virol. 1975 May;15(5):1239–1247. doi: 10.1128/jvi.15.5.1239-1247.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Finch J. T., Klug A. The structure of viruses of the papilloma-polyoma type 3. Structure of rabbit papilloma virus, with an appendix on the topography of contrast in negative-staining for electron-microscopy. J Mol Biol. 1965 Aug;13(1):1–12. doi: 10.1016/s0022-2836(65)80075-4. [DOI] [PubMed] [Google Scholar]
  11. Firzlaff J. M., Kiviat N. B., Beckmann A. M., Jenison S. A., Galloway D. A. Detection of human papillomavirus capsid antigens in various squamous epithelial lesions using antibodies directed against the L1 and L2 open reading frames. Virology. 1988 Jun;164(2):467–477. doi: 10.1016/0042-6822(88)90561-2. [DOI] [PubMed] [Google Scholar]
  12. Galloway D. A., McDougall J. K. Human papillomaviruses and carcinomas. Adv Virus Res. 1989;37:125–171. doi: 10.1016/s0065-3527(08)60834-9. [DOI] [PubMed] [Google Scholar]
  13. Griffith J. P., Griffith D. L., Rayment I., Murakami W. T., Caspar D. L. Inside polyomavirus at 25-A resolution. Nature. 1992 Feb 13;355(6361):652–654. doi: 10.1038/355652a0. [DOI] [PubMed] [Google Scholar]
  14. Haffar O., Garrigues J., Travis B., Moran P., Zarling J., Hu S. L. Human immunodeficiency virus-like, nonreplicating, gag-env particles assemble in a recombinant vaccinia virus expression system. J Virol. 1990 Jun;64(6):2653–2659. doi: 10.1128/jvi.64.6.2653-2659.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hu S. L., Travis B. M., Garrigues J., Zarling J. M., Sridhar P., Dykers T., Eichberg J. W., Alpers C. Processing, assembly, and immunogenicity of human immunodeficiency virus core antigens expressed by recombinant vaccinia virus. Virology. 1990 Nov;179(1):321–329. doi: 10.1016/0042-6822(90)90300-g. [DOI] [PubMed] [Google Scholar]
  16. Jenison S. A., Firzlaff J. M., Langenberg A., Galloway D. A. Identification of immunoreactive antigens of human papillomavirus type 6b by using Escherichia coli-expressed fusion proteins. J Virol. 1988 Jun;62(6):2115–2123. doi: 10.1128/jvi.62.6.2115-2123.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Jenison S. A., Yu X. P., Valentine J. M., Galloway D. A. Human antibodies react with an epitope of the human papillomavirus type 6b L1 open reading frame which is distinct from the type-common epitope. J Virol. 1989 Feb;63(2):809–818. doi: 10.1128/jvi.63.2.809-818.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Koerner T. J., Hill J. E., Myers A. M., Tzagoloff A. High-expression vectors with multiple cloning sites for construction of trpE fusion genes: pATH vectors. Methods Enzymol. 1991;194:477–490. doi: 10.1016/0076-6879(91)94036-c. [DOI] [PubMed] [Google Scholar]
  19. Komly C. A., Breitburd F., Croissant O., Streeck R. E. The L2 open reading frame of human papillomavirus type 1a encodes a minor structural protein carrying type-specific antigens. J Virol. 1986 Nov;60(2):813–816. doi: 10.1128/jvi.60.2.813-816.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kopan R., Traska G., Fuchs E. Retinoids as important regulators of terminal differentiation: examining keratin expression in individual epidermal cells at various stages of keratinization. J Cell Biol. 1987 Jul;105(1):427–440. doi: 10.1083/jcb.105.1.427. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Koutsky L. A., Galloway D. A., Holmes K. K. Epidemiology of genital human papillomavirus infection. Epidemiol Rev. 1988;10:122–163. doi: 10.1093/oxfordjournals.epirev.a036020. [DOI] [PubMed] [Google Scholar]
  22. Kreider J. W., Howett M. K., Leure-Dupree A. E., Zaino R. J., Weber J. A. Laboratory production in vivo of infectious human papillomavirus type 11. J Virol. 1987 Feb;61(2):590–593. doi: 10.1128/jvi.61.2.590-593.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kreider J. W., Patrick S. D., Cladel N. M., Welsh P. A. Experimental infection with human papillomavirus type 1 of human hand and foot skin. Virology. 1990 Jul;177(1):415–417. doi: 10.1016/0042-6822(90)90503-j. [DOI] [PubMed] [Google Scholar]
  24. Liddington R. C., Yan Y., Moulai J., Sahli R., Benjamin T. L., Harrison S. C. Structure of simian virus 40 at 3.8-A resolution. Nature. 1991 Nov 28;354(6351):278–284. doi: 10.1038/354278a0. [DOI] [PubMed] [Google Scholar]
  25. Mackett M., Smith G. L., Moss B. General method for production and selection of infectious vaccinia virus recombinants expressing foreign genes. J Virol. 1984 Mar;49(3):857–864. doi: 10.1128/jvi.49.3.857-864.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Orth G., Favre M., Croissant O. Characterization of a new type of human papillomavirus that causes skin warts. J Virol. 1977 Oct;24(1):108–120. doi: 10.1128/jvi.24.1.108-120.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Piccini A., Paoletti E. Vaccinia: virus, vector, vaccine. Adv Virus Res. 1988;34:43–64. doi: 10.1016/s0065-3527(08)60515-1. [DOI] [PubMed] [Google Scholar]
  28. Rayment I., Baker T. S., Caspar D. L., Murakami W. T. Polyoma virus capsid structure at 22.5 A resolution. Nature. 1982 Jan 14;295(5845):110–115. doi: 10.1038/295110a0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Salunke D. M., Caspar D. L., Garcea R. L. Self-assembly of purified polyomavirus capsid protein VP1. Cell. 1986 Sep 12;46(6):895–904. doi: 10.1016/0092-8674(86)90071-1. [DOI] [PubMed] [Google Scholar]
  30. Tomita Y., Shirasawa H., Sekine H., Simizu B. Expression of the human papillomavirus type 6b L2 open reading frame in Escherichia coli: L2-beta-galactosidase fusion proteins and their antigenic properties. Virology. 1987 May;158(1):8–14. doi: 10.1016/0042-6822(87)90231-5. [DOI] [PubMed] [Google Scholar]
  31. Tomita Y., Shirasawa H., Simizu B. Expression of human papillomavirus types 6b and 16 L1 open reading frames in Escherichia coli: detection of a 56,000-dalton polypeptide containing genus-specific (common) antigens. J Virol. 1987 Aug;61(8):2389–2394. doi: 10.1128/jvi.61.8.2389-2394.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Watt F. M., Green H. Stratification and terminal differentiation of cultured epidermal cells. Nature. 1982 Feb 4;295(5848):434–436. doi: 10.1038/295434a0. [DOI] [PubMed] [Google Scholar]
  33. Winokur P. L., McLinden J. H., Stapleton J. T. The hepatitis A virus polyprotein expressed by a recombinant vaccinia virus undergoes proteolytic processing and assembly into viruslike particles. J Virol. 1991 Sep;65(9):5029–5036. doi: 10.1128/jvi.65.9.5029-5036.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Yaegashi N., Jenison S. A., Valentine J. M., Dunn M., Taichman L. B., Baker D. A., Galloway D. A. Characterization of murine polyclonal antisera and monoclonal antibodies generated against intact and denatured human papillomavirus type 1 virions. J Virol. 1991 Mar;65(3):1578–1583. doi: 10.1128/jvi.65.3.1578-1583.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Zhou J., Doorbar J., Sun X. Y., Crawford L. V., McLean C. S., Frazer I. H. Identification of the nuclear localization signal of human papillomavirus type 16 L1 protein. Virology. 1991 Dec;185(2):625–632. doi: 10.1016/0042-6822(91)90533-h. [DOI] [PubMed] [Google Scholar]
  36. Zhou J., Sun X. Y., Stenzel D. J., Frazer I. H. Expression of vaccinia recombinant HPV 16 L1 and L2 ORF proteins in epithelial cells is sufficient for assembly of HPV virion-like particles. Virology. 1991 Nov;185(1):251–257. doi: 10.1016/0042-6822(91)90772-4. [DOI] [PubMed] [Google Scholar]
  37. zur Hausen H. Human papillomaviruses in the pathogenesis of anogenital cancer. Virology. 1991 Sep;184(1):9–13. doi: 10.1016/0042-6822(91)90816-t. [DOI] [PubMed] [Google Scholar]

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

RESOURCES