1887

Abstract

The human papillomavirus (HPV) type 16 major capsid proteins L1 and L2 have been produced in a baculovirus expression system. Both proteins are expressed at a high level and can be readily solubilized. The L1 capsid protein migrates close to its expected of 60K. On the other hand L2 exhibits a much higher migrating at 73K, which is considerably greater than its predicted of 50K. The identity of both proteins has been confirmed also by Western blot analysis. Both proteins are produced in drastically reduced amounts in the presence of tunicamycin. In addition both L1 and L2 show interesting patterns of phosphorylation. L1 is phosphorylated only weakly and this appears to be quite labile, whereas L2 is very heavily phosphorylated and this, in contrast, appears to be very stable. We have also made use of a dual expression vector for co-expressing the L1 and L2 proteins within the same baculovirus-infected cell. The results obtained from this system demonstrate the presence of protein complexes forming between the two capsid proteins. These studies indicate that at least the initial events in capsid assembly of HPVs can occur in the absence of viral DNA.

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1991-12-01
2024-12-06
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References

  1. Banks L., Matlashewski G., Pim D., Churcher M., Roberts C., Crawford L. 1987; Expression of human papillomavirus type 6 and 16 capsid proteins in bacteria and their antigenic characterization. Journal of General Virology 68:3081–3089
    [Google Scholar]
  2. Browne H. M., Churcher M. J., Stanley M. A., Smith G. L., Minson A. C. 1988; Analysis of the L1 gene product of human papillomavirus type 16 by expression in a vaccinia virus recombinant. Journal of General Virology 69:1263–1273
    [Google Scholar]
  3. Doorbar J., Gallimore P. H. 1987; Identification of proteins encoded by the L1 and L2 open reading frames of human papillomavirus la. Journal of Virology 61:2793–2799
    [Google Scholar]
  4. Evan G. I., Lewis G. K., Ramsey G., Bishop J. M. 1985; Isolation of monoclonal antibodies specific for human c-myc proto oncogene product. Molecular and Cellular Biology 5:3610–3616
    [Google Scholar]
  5. Firzlaff J. M., Kiviat N. B., Beckmann A. M., Jenison S. A., Galloway D. A. 1987; Detection of human papillomavirus capsid antigens in various squamous epithelial lesions using antibodies directed against the L1 and L2 open reading frames. Virology 164:467–477
    [Google Scholar]
  6. French T. J., Roy P. 1990; Synthesis of bluetongue virus (BTV) corelike particles by a recombinant baculovirus expressing the two major structural core proteins of BTV. Journal of Virology 64:1530–1536
    [Google Scholar]
  7. Hawley-Nelson P., Vousden K. H., Hubbert N. L., Lowy D. R., Schiller J. T. 1989; HPV E6 and E7 proteins cooperate to immortalise human foreskin keratinocytes. EMBO Journal 8:3905–3910
    [Google Scholar]
  8. Komly C. A., Breitburd F., Croissant O., Streeck R. E. 1986; The L2 open reading frame of human papillomavirus type la encodes a minor structural protein carrying type specific antigens. Journal of Virology 60:813–816
    [Google Scholar]
  9. MacLean C. F., Churcher M. J., Meinke J., Smith G. L., Higgins G., Stanley M., Minson A. C. 1990; Production and characterisation of a monoclonal antibody to human papillomavirus type 16 using a recombinant vaccinia virus. Journal of Clinical Pathology 43:488–492
    [Google Scholar]
  10. Matlashewski G. 1989; The cell biology of human papillomavirus transformed cells. Anticancer Research 9:1447–1556
    [Google Scholar]
  11. Matlashewski G., Schneider J., Banks L., Jones N., Murray A., Crawford L. 1987; Human papillomavirus type 16 DNA cooperates with activated ras in transforming cells. EMBO Journal 6:1741–1746
    [Google Scholar]
  12. Munger K., Phelps W. C., Bubb V., Howley P. M., Schlegel R. 1989; The E6 and E7 genes of the human papillomavirus type 16 are together necessary and sufficient for transformation of primary human keratinocytes. Journal of Virology 63:4417–4421
    [Google Scholar]
  13. Page M. J. 1989; p36C: an improved baculovirus vector for producing high levels of mature recombinant proteins. Nucleic Acids Research 17:454
    [Google Scholar]
  14. Parton A. 1990; Nucleotide sequence of the HPV-16 L1 open reading frame. Nucleic Acids Research 18:3631
    [Google Scholar]
  15. Phelps W. C., Yee C. L., Munger K., Howley P. M. 1988; The human papillomavirus type 16 E7 gene encodes transactivation and transformation functions similar to those of adenovirus ElA. Cell 53:539–547
    [Google Scholar]
  16. Rippe R., Meinke W. 1989; Identification and characterisation of the BPV-2 L2 protein. Virology 171:298–301
    [Google Scholar]
  17. Rose R. C., Bonnez W., Strike D. G., Reichman R. C. 1990; Expression of the full-length products of the human papillomavirus type 6b (HPV-6b) and HPV-11 L2 open reading frames by recombinant baculovirus, and antigenic comparisons with HPV-11 whole virus particles. Journal of General Virology 71:2725–2729
    [Google Scholar]
  18. Storey A., Pim D., Murray A., Osborn K., Banks L., Crawford L. 1988; Comparison of the in vitro transforming activities of human papillomavirus types. EMBO Journal 7:1815–1820
    [Google Scholar]
  19. Vousden K. 1989; Human papillomaviruses and cervical carcinoma. Cancer Cells 1:43–50
    [Google Scholar]
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