1887

Abstract

The E7 open reading frame of human papillomavirus type 16 (HPV-16) encodes a protein that can immortalize primary rat cells, cooperate with the oncoprotein to transform low passage rat cells and transform established rodent cells to anchorage independence. The immortalizing and cooperation functions have been investigated using a series of point mutations that introduce single amino acid changes into the E7 protein in two distinct regions. Certain mutations altering amino acids conserved between the E7 protein of genital HPV types, the adenovirus E1 a protein and simian virus 40 large T antigen abolished the ability of the E7 protein to immortalize or cooperate with in a focus forming assay. Mutations in a consensus sequence for a casein kinase II recognition site, which is also shared by E1a and large T, reduced immortalizing activity, but did not affect the ability to cooperate with . Single mutations disrupting cysteine motifs, which form putative zinc-binding sites in the second region, reduced the activity of the E7 protein, whereas double mutants, in which neither of the cysteine motifs remained intact, showed no or very low activity. The activity of the mutants in immortalization and cooperation assays was essentially the same as their transforming activities in NIH 3T3 cells. This indicates that these three functions of E7 map to overlapping domains which cannot be separated by these mutations in the region of E1a/large T homology or the cysteine motifs.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-71-2-449
1990-02-01
2024-03-28
Loading full text...

Full text loading...

/deliver/fulltext/jgv/71/2/JV0710020449.html?itemId=/content/journal/jgv/10.1099/0022-1317-71-2-449&mimeType=html&fmt=ahah

References

  1. Barbosa M. S., Lowy D. R., Schiller J. T. 1989; Papillomavirus polypeptides E6 and E7 are zinc-binding proteins. Journal of Virology 63:1404–1407
    [Google Scholar]
  2. Chesters P. M., Mccance D. J. 1989; Human papillomavirus types 6 and 16 in cooperation with Ha-rastransform secondary rat embryo fibroblasts. Journal of General Virology 70:353–365
    [Google Scholar]
  3. Cole S. T., Danos O. 1987; Nucleotide sequence and comparative analysis of the human papillomavirus type 18 genome: phylogeny of papillomaviruses and repeated structure of the E6 and E7 gene products. Journal of Molecular Biology 193:599–608
    [Google Scholar]
  4. Crook T., Storey A., Almond N., Osborn K., Crawford L. 1988; Human papillomavirus type 16 cooperates with activated ras and fos oncogenes in the hormone-dependent transformation of primary mouse cells. Proceedings of the National Academy of Sciences, U.S.A 85:8820–8824
    [Google Scholar]
  5. De Caprio J. A., Ludlow J. W., Figge J., Shew J.-Y., Huang C.-M., Lee W.-H., Masilio E., Paucha E., Livingston D. M. 1988; SV40 large tumor antigen forms a specific complex with the product of the retinoblastoma susceptibility gene. Cell 54:275–283
    [Google Scholar]
  6. Dürst M., Gissmann L., Ikenberg H., Zur Hausen H. 1983; A papillomavirus DNA from a cervical carcinoma and its prevalence in cancer biopsy samples from different geographic regions. Proceedings of the National Academy of Sciences, U.S.A 80:3812–3815
    [Google Scholar]
  7. Dyson N., Howley P. M., Munger K., Harlow E. 1989; The human papillomavirus-16 E7 oncoprotein is able to bind to the retinoblastoma gene product. Science 243:934–937
    [Google Scholar]
  8. Edmonds C., Vousden K. H. 1989; A point mutational analysis of human papillomavirus type 16 E7 protein. Journal of Virology 63:2650–2656
    [Google Scholar]
  9. Evans R. M., Hollenberg S. M. 1988; Zinc fingers: gilt by association. Cell 53:675
    [Google Scholar]
  10. Gissmann L., Boshart M., Durst M., Ikenberg H., Wagner D. 1984; Presence of human papillomavirus (HPV) DNA in genital tumours. Journal of Investigative Dermatology 83:265–288
    [Google Scholar]
  11. Jat P. S., Sharp P. A. 1989; Cell lines established by a temperature sensitive SV40 large T antigen are growth restricted at the nonpermissive temperature. Molecular and Cellular Biology 9:1672–1681
    [Google Scholar]
  12. Kalderon D., Smith A. E. 1984; In vitro mutagenesis of a putative DNA binding domain of SV40 large-T. Virology 139:109–137
    [Google Scholar]
  13. Kanda T., Watanabe S., Yoshiike K. 1988; Immortalisation of primary rat cells by human papillomavirus type 16 subgenomic DNA fragments controlled by the SV40 promoter. Virology 165:321–325
    [Google Scholar]
  14. Kuppuswamy M., Subramanian T., Chinnadurai G. 1988; Separation of immortalization and T24-ras oncogene cooperative functions of adenovirus E1a. Oncogene 2:613–615
    [Google Scholar]
  15. Lancaster W. D., Olson C. 1982; Animal papillomaviruses. Microbiological Reviews 46:191–207
    [Google Scholar]
  16. Land H., Parada L. F., Weinberg R. 1983; Tumorigenic conversion of primary embryo fibroblasts requires at least two cooperating oncogenes. Nature; London: 304596–602
    [Google Scholar]
  17. Lillie J. W., Green M., Green M. R. 1986; An adenovirus E1a protein region required for transformation and transcriptional repression. Cell 46:1043–1051
    [Google Scholar]
  18. Moran E. 1988; A region of SV40 large T antigen can substitute for a transforming domain of the adenovirus E1a products. Nature; London: 334168–170
    [Google Scholar]
  19. Moran E., Zerler B., Harrison T. M., Mathews M. B. 1986; Identification of separate domains in the adenovirus E1A gene for immortalisation activity and the activation of virus early genes. Molecular and Cellular Biology 6:3470–3480
    [Google Scholar]
  20. Pfister H. 1984; Biology and biochemistry of papillomaviruses. Reviews of Physiology, Biochemistry and Pharmacology 99:111–181
    [Google Scholar]
  21. 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 E1a. Cell 53:539–547
    [Google Scholar]
  22. Smith D. H., Ziff E. B. 1988; The amino-terminal region of the adenovirus serotype 5 Elaprotein performs two separate functions when expressed in primary rat kidney cells. Molecular and Cellular Biology 8:3882–3890
    [Google Scholar]
  23. Smotkin D., Wettstein F. O. 1987; The major human papillomavirus protein in cervical cancers is a cytoplasmic phosphoprotein. Journal of Virology 61:1686–1689
    [Google Scholar]
  24. 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 1:1815–1820
    [Google Scholar]
  25. Sugden B., Marsh K., Yates J. 1985; A vector that replicates as a plasmid and can be efficiently selected in B-lymphoblasts transformed by Epstein-Barr virus. Molecular and Cellular Biology 5:410–413
    [Google Scholar]
  26. Vousden K. H., Jat P. S. 1989; Functional similarity between HPV 16 E7, SV40 large T and adenovirus E1a proteins. Oncogene 5:153–158
    [Google Scholar]
  27. Vousden K. H., Doniger J., Dipaolo J. A., Lowy D. R. 1988; The E7 open reading frame of human papillomavirus type 16 encodes a transforming gene. Oncogene Research 3:167–175
    [Google Scholar]
  28. Whyte P., Ruley H. E., Harlow E. 1988; Two regions of the adenovirus early region 1A protein are required for transformation. Journal of Virology 62:257–265
    [Google Scholar]
  29. Zur Hausen H. 1989; Papillomavirus as carcinomaviruses. In Advances in Viral Oncology 8 pp. 1–26 Klein G. Edited by New York: Raven Press;
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-71-2-449
Loading
/content/journal/jgv/10.1099/0022-1317-71-2-449
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error