Skip to main content
Journal of Virology logoLink to Journal of Virology
. 1995 Jul;69(7):4489–4494. doi: 10.1128/jvi.69.7.4489-4494.1995

Human papillomavirus type 16 E5 protein affects cell-cell communication in an epithelial cell line.

I Oelze 1, J Kartenbeck 1, K Crusius 1, A Alonso 1
PMCID: PMC189191  PMID: 7769709

Abstract

The human papillomavirus type 16 (HPV16) E5 protein is considered to have weak oncogenic properties, and its function in infected human keratinocytes is unknown. HPV16 E5 protein has been found to localize to the Golgi apparatus and the plasma membrane. To analyze the effect of E5 on plasma membrane properties, cells from the human keratinocyte cell line HaCaT were transfected with the HPV16 E5 open reading frame under the control of an inducible promoter. The gap junction-mediated cell-cell communication of E5- and vector-transfected cells was analyzed by microinjection of Lucifer yellow to measure dye coupling of the cells. A strong impairment of dye transfer in E5-transfected cells but not in vector-transfected cells was observed, with more than 80% dye transfer inhibition 40 min after injection. This impairment correlated with dephosphorylation of connexin 43, the major gap junctional protein in HaCaT cells. Furthermore, the dye coupling inhibition was not the result of differentiation of the E5-expressing cells, since no overexpression of cytokeratin 1 or filaggrin, markers of HaCaT cell differentiation, could be observed. These results therefore strongly suggest a correlation between expression of the HPV16 E5 open reading frame, impairment of gap junction-mediated dye coupling, and dephosphorylation of connexin 43.

Full Text

The Full Text of this article is available as a PDF (960.7 KB).

Selected References

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

  1. Boukamp P., Petrussevska R. T., Breitkreutz D., Hornung J., Markham A., Fusenig N. E. Normal keratinization in a spontaneously immortalized aneuploid human keratinocyte cell line. J Cell Biol. 1988 Mar;106(3):761–771. doi: 10.1083/jcb.106.3.761. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bouvard V., Matlashewski G., Gu Z. M., Storey A., Banks L. The human papillomavirus type 16 E5 gene cooperates with the E7 gene to stimulate proliferation of primary cells and increases viral gene expression. Virology. 1994 Aug 15;203(1):73–80. doi: 10.1006/viro.1994.1456. [DOI] [PubMed] [Google Scholar]
  3. Breitkreutz D., Stark H. J., Plein P., Baur M., Fusenig N. E. Differential modulation of epidermal keratinization in immortalized (HaCaT) and tumorigenic human skin keratinocytes (HaCaT-ras) by retinoic acid and extracellular Ca2+. Differentiation. 1993 Oct;54(3):201–217. doi: 10.1111/j.1432-0436.1993.tb01602.x. [DOI] [PubMed] [Google Scholar]
  4. Brissette J. L., Kumar N. M., Gilula N. B., Hall J. E., Dotto G. P. Switch in gap junction protein expression is associated with selective changes in junctional permeability during keratinocyte differentiation. Proc Natl Acad Sci U S A. 1994 Jul 5;91(14):6453–6457. doi: 10.1073/pnas.91.14.6453. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Burkhardt A., Willingham M., Gay C., Jeang K. T., Schlegel R. The E5 oncoprotein of bovine papillomavirus is oriented asymmetrically in Golgi and plasma membranes. Virology. 1989 May;170(1):334–339. doi: 10.1016/0042-6822(89)90391-7. [DOI] [PubMed] [Google Scholar]
  6. Cohen B. D., Goldstein D. J., Rutledge L., Vass W. C., Lowy D. R., Schlegel R., Schiller J. T. Transformation-specific interaction of the bovine papillomavirus E5 oncoprotein with the platelet-derived growth factor receptor transmembrane domain and the epidermal growth factor receptor cytoplasmic domain. J Virol. 1993 Sep;67(9):5303–5311. doi: 10.1128/jvi.67.9.5303-5311.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Conrad M., Bubb V. J., Schlegel R. The human papillomavirus type 6 and 16 E5 proteins are membrane-associated proteins which associate with the 16-kilodalton pore-forming protein. J Virol. 1993 Oct;67(10):6170–6178. doi: 10.1128/jvi.67.10.6170-6178.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Conrad M., Goldstein D., Andresson T., Schlegel R. The E5 protein of HPV-6, but not HPV-16, associates efficiently with cellular growth factor receptors. Virology. 1994 May 1;200(2):796–800. doi: 10.1006/viro.1994.1244. [DOI] [PubMed] [Google Scholar]
  9. Dyson N., Howley P. M., Münger K., Harlow E. The human papilloma virus-16 E7 oncoprotein is able to bind to the retinoblastoma gene product. Science. 1989 Feb 17;243(4893):934–937. doi: 10.1126/science.2537532. [DOI] [PubMed] [Google Scholar]
  10. Dürst M., Dzarlieva-Petrusevska R. T., Boukamp P., Fusenig N. E., Gissmann L. Molecular and cytogenetic analysis of immortalized human primary keratinocytes obtained after transfection with human papillomavirus type 16 DNA. Oncogene. 1987;1(3):251–256. [PubMed] [Google Scholar]
  11. Finbow M. E., Pitts J. D. Is the gap junction channel--the connexon--made of connexin or ductin? J Cell Sci. 1993 Oct;106(Pt 2):463–471. doi: 10.1242/jcs.106.2.463. [DOI] [PubMed] [Google Scholar]
  12. Fitzgerald D. J., Fusenig N. E., Boukamp P., Piccoli C., Mesnil M., Yamasaki H. Expression and function of connexin in normal and transformed human keratinocytes in culture. Carcinogenesis. 1994 Sep;15(9):1859–1865. doi: 10.1093/carcin/15.9.1859. [DOI] [PubMed] [Google Scholar]
  13. Goldstein D. J., Li W., Wang L. M., Heidaran M. A., Aaronson S., Shinn R., Schlegel R., Pierce J. H. The bovine papillomavirus type 1 E5 transforming protein specifically binds and activates the beta-type receptor for the platelet-derived growth factor but not other related tyrosine kinase-containing receptors to induce cellular transformation. J Virol. 1994 Jul;68(7):4432–4441. doi: 10.1128/jvi.68.7.4432-4441.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kamibayashi Y., Oyamada M., Oyamada Y., Mori M. Expression of gap junction proteins connexin 26 and 43 is modulated during differentiation of keratinocytes in newborn mouse epidermis. J Invest Dermatol. 1993 Dec;101(6):773–778. doi: 10.1111/1523-1747.ep12371693. [DOI] [PubMed] [Google Scholar]
  15. Kell B., Jewers R. J., Cason J., Pakarian F., Kaye J. N., Best J. M. Detection of E5 oncoprotein in human papillomavirus type 16-positive cervical scrapes using antibodies raised to synthetic peptides. J Gen Virol. 1994 Sep;75(Pt 9):2451–2456. doi: 10.1099/0022-1317-75-9-2451. [DOI] [PubMed] [Google Scholar]
  16. Leechanachai P., Banks L., Moreau F., Matlashewski G. The E5 gene from human papillomavirus type 16 is an oncogene which enhances growth factor-mediated signal transduction to the nucleus. Oncogene. 1992 Jan;7(1):19–25. [PubMed] [Google Scholar]
  17. Leptak C., Ramon y Cajal S., Kulke R., Horwitz B. H., Riese D. J., 2nd, Dotto G. P., DiMaio D. Tumorigenic transformation of murine keratinocytes by the E5 genes of bovine papillomavirus type 1 and human papillomavirus type 16. J Virol. 1991 Dec;65(12):7078–7083. doi: 10.1128/jvi.65.12.7078-7083.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Matesic D. F., Rupp H. L., Bonney W. J., Ruch R. J., Trosko J. E. Changes in gap-junction permeability, phosphorylation, and number mediated by phorbol ester and non-phorbol-ester tumor promoters in rat liver epithelial cells. Mol Carcinog. 1994 Aug;10(4):226–236. doi: 10.1002/mc.2940100407. [DOI] [PubMed] [Google Scholar]
  19. Oyamada M., Kimura H., Oyamada Y., Miyamoto A., Ohshika H., Mori M. The expression, phosphorylation, and localization of connexin 43 and gap-junctional intercellular communication during the establishment of a synchronized contraction of cultured neonatal rat cardiac myocytes. Exp Cell Res. 1994 Jun;212(2):351–358. doi: 10.1006/excr.1994.1154. [DOI] [PubMed] [Google Scholar]
  20. Petti L., DiMaio D. Specific interaction between the bovine papillomavirus E5 transforming protein and the beta receptor for platelet-derived growth factor in stably transformed and acutely transfected cells. J Virol. 1994 Jun;68(6):3582–3592. doi: 10.1128/jvi.68.6.3582-3592.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Pim D., Collins M., Banks L. Human papillomavirus type 16 E5 gene stimulates the transforming activity of the epidermal growth factor receptor. Oncogene. 1992 Jan;7(1):27–32. [PubMed] [Google Scholar]
  22. Pirisi L., Yasumoto S., Feller M., Doniger J., DiPaolo J. A. Transformation of human fibroblasts and keratinocytes with human papillomavirus type 16 DNA. J Virol. 1987 Apr;61(4):1061–1066. doi: 10.1128/jvi.61.4.1061-1066.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Scheffner M., Werness B. A., Huibregtse J. M., Levine A. J., Howley P. M. The E6 oncoprotein encoded by human papillomavirus types 16 and 18 promotes the degradation of p53. Cell. 1990 Dec 21;63(6):1129–1136. doi: 10.1016/0092-8674(90)90409-8. [DOI] [PubMed] [Google Scholar]
  24. Schmelz M., Duden R., Cowin P., Franke W. W. A constitutive transmembrane glycoprotein of Mr 165,000 (desmoglein) in epidermal and non-epidermal desmosomes. II. Immunolocalization and microinjection studies. Eur J Cell Biol. 1986 Dec;42(2):184–199. [PubMed] [Google Scholar]
  25. Stoler M. H., Rhodes C. R., Whitbeck A., Wolinsky S. M., Chow L. T., Broker T. R. Human papillomavirus type 16 and 18 gene expression in cervical neoplasias. Hum Pathol. 1992 Feb;23(2):117–128. doi: 10.1016/0046-8177(92)90232-r. [DOI] [PubMed] [Google Scholar]
  26. Straight S. W., Hinkle P. M., Jewers R. J., McCance D. J. The E5 oncoprotein of human papillomavirus type 16 transforms fibroblasts and effects the downregulation of the epidermal growth factor receptor in keratinocytes. J Virol. 1993 Aug;67(8):4521–4532. doi: 10.1128/jvi.67.8.4521-4532.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Willecke K., Hennemann H., Dahl E., Jungbluth S., Heynkes R. The diversity of connexin genes encoding gap junctional proteins. Eur J Cell Biol. 1991 Oct;56(1):1–7. [PubMed] [Google Scholar]
  28. 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