1887

Journal of General Virology header logo

Volume 97, Issue 2

Disc large 1 expression is altered by human papillomavirus E6/E7 proteins in organotypic cultures of human keratinocytes Free

Abstract

Loss of cell polarity is a fundamental process in cell transformation. Among polarity proteins, we focused on human disc large (DLG1), which is localized mainly at adherens junctions and contributes to the control of cell proliferation. We previously demonstrated that its expression is altered in HPV-associated cervical neoplastic lesions, but the mechanisms beyond this remain unknown. In this study, we analysed the contribution of HPV proteins to the changes in DLG1 expression in the squamous epithelium. We observed tissue and intracellular misdistribution of DLG1 when high-risk HPV-18 E7 or E6/E7 proteins were expressed in organotypic raft cultures. The viral oncoproteins induce the loss of DLG1 from the cell borders and an increase in the level of DLG1 protein, reflecting the pattern observed in cervical lesions. These findings were corroborated in cultures bearing the entire HPV-18 genome. Interestingly, changes in tissue distribution and abundance of DLG1 were also detected in organotypic cultures expressing the low-risk HPV-11 E7 or E6/E7 proteins, suggesting a conserved function among different HPV types. However, for low-risk HPVs, the subcellular localization of DLG1 at cell-to-cell contacts was predominantly maintained. This report offers new evidence, we believe, of the involvement of HPV proteins in DLG1 expression pattern and our data support previous observations regarding DLG1 expression in cervical lesions.

  • Received:
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  • Published Online:
© 2015 The Authors
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2016-02-01
2024-04-20
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References

  1. Assémat E., Bazellières E., Pallesi-Pocachard E., Le Bivic A., Massey-Harroche D. 2008; Polarity complex proteins. Biochim Biophys Acta 1778:614–630 [View Article][PubMed]
     [Google Scholar]
  2. Bilder D. 2004; Epithelial polarity and proliferation control: links from the Drosophila neoplastic tumor suppressors. Genes Dev 18:1909–1925 [View Article][PubMed]
     [Google Scholar]
  3. Boccardo E., Noya F., Broker T. R., Chow L. T., Villa L. L. 2004; HPV-18 confers resistance to TNF-α in organotypic cultures of human keratinocytes. Virology 328:233–243 [View Article][PubMed]
     [Google Scholar]
  4. Boccardo E., Manzini Baldi C. V., Carvalho A. F., Rabachini T., Torres C., Barreta L. A., Brentani H., Villa L. L. 2010; Expression of human papillomavirus type 16 E7 oncoprotein alters keratinocytes expression profile in response to tumor necrosis factor-α. Carcinogenesis 31:521–531 [View Article][PubMed]
     [Google Scholar]
  5. Caberg J. H., Hubert P. M., Begon D. Y., Herfs M. F., Roncarati P. J., Boniver J. J., Delvenne P. O. 2008; Silencing of E7 oncogene restores functional E-cadherin expression in human papillomavirus 16-transformed keratinocytes. Carcinogenesis 29:1441–1447 [View Article][PubMed]
     [Google Scholar]
  6. Cavatorta A. L., Fumero G., Chouhy D., Aguirre R., Nocito A. L., Giri A. A., Banks L., Gardiol D. 2004; Differential expression of the human homologue of Drosophila discs large oncosuppressor in histologic samples from human papillomavirus-associated lesions as a marker for progression to malignancy. Int J Cancer 111:373–380 [View Article][PubMed]
     [Google Scholar]
  7. Cheng S., Schmidt-Grimminger D.-C., Murant T., Broker T. R., Chow L. T. 1995; Differentiation-dependent up-regulation of the human papillomavirus E7 gene reactivates cellular DNA replication in suprabasal differentiated keratinocytes. Genes Dev 9:2335–2349 [View Article][PubMed]
     [Google Scholar]
  8. Delury C. P., Marsh E. K., James C. D., Boon S. S., Banks L., Knight G. L., Roberts S. 2013; The role of protein kinase A regulation of the E6 PDZ-binding domain during the differentiation-dependent life cycle of human papillomavirus type 18. J Virol 87:9463–9472 [View Article][PubMed]
     [Google Scholar]
  9. Facciuto F., Cavatorta A. L., Valdano M. B., Marziali F., Gardiol D. 2012; Differential expression of PDZ domain-containing proteins in human diseases - challenging topics and novel issues. FEBS J 279:3538–3548 [View Article][PubMed]
     [Google Scholar]
  10. Facciuto F., Bugnon Valdano M., Marziali F., Massimi P., Banks L., Cavatorta A. L., Gardiol D. 2014; Human papillomavirus (HPV)-18 E6 oncoprotein interferes with the epithelial cell polarity Par3 protein. Mol Oncol 8:533–543 [View Article][PubMed]
     [Google Scholar]
  11. Fang L., Meyers C., Budgeon L. R., Howett M. K. 2006; Induction of productive human papillomavirus type 11 life cycle in epithelial cells grown in organotypic raft cultures. Virology 347:28–35 [View Article][PubMed]
     [Google Scholar]
  12. Flores E. R., Allen-Hoffmann B. L., Lee D., Lambert P. F. 2000; The human papillomavirus type 16 E7 oncogene is required for the productive stage of the viral life cycle. J Virol 74:6622–6631 [View Article][PubMed]
     [Google Scholar]
  13. Frese K. K., Latorre I. J., Chung S. H., Caruana G., Bernstein A., Jones S. N., Donehower L. A., Justice M. J., Garner C. C., Javier R. T. 2006; Oncogenic function for the Dlg1 mammalian homolog of the Drosophila discs-large tumor suppressor. EMBO J 25:1406–1417 [View Article][PubMed]
     [Google Scholar]
  14. Fuja T. J., Lin F., Osann K. E., Bryant P. J. 2004; Somatic mutations and altered expression of the candidate tumor suppressors CSNK1ϵ, DLG1, and EDD/hHYD in mammary ductal carcinoma. Cancer Res 64:942–951 [View Article][PubMed]
     [Google Scholar]
  15. Gardiol D., Kühne C., Glaunsinger B., Lee S. S., Javier R., Banks L. 1999; Oncogenic human papillomavirus E6 proteins target the discs large tumour suppressor for proteasome-mediated degradation. Oncogene 18:5487–5496 [View Article][PubMed]
     [Google Scholar]
  16. Gardiol D., Zacchi A., Petrera F., Stanta G., Banks L. 2006; Human discs large and scrib are localized at the same regions in colon mucosa and changes in their expression patterns are correlated with loss of tissue architecture during malignant progression. Int J Cancer 119:1285–1290 [View Article][PubMed]
     [Google Scholar]
  17. Gaudet S., Branton D., Lue R. A. 2000; Characterization of PDZ-binding kinase, a mitotic kinase. Proc Natl Acad Sci U S A 97:5167–5172 [View Article][PubMed]
     [Google Scholar]
  18. Gaudet S., Langlois M. J., Lue R. A., Rivard N., Viel A. 2011; The MEK2-binding tumor suppressor hDlg is recruited by E-cadherin to the midbody ring. BMC Cell Biol 12:55 [View Article][PubMed]
     [Google Scholar]
  19. Guess J. C., McCance D. J. 2005; Decreased migration of Langerhans precursor-like cells in response to human keratinocytes expressing human papillomavirus type 16 E6/E7 is related to reduced macrophage inflammatory protein-3α production. J Virol 79:14852–14862 [View Article][PubMed]
     [Google Scholar]
  20. Gyöngyösi E., Szalmás A., Ferenczi A., Kónya J., Gergely L., Veress G. 2012; Effects of human papillomavirus (HPV) type 16 oncoproteins on the expression of involucrin in human keratinocytes. Virol J 9:36 [View Article][PubMed]
     [Google Scholar]
  21. Hellner K., Mar J., Fang F., Quackenbush J., Münger K. 2009; HPV16 E7 oncogene expression in normal human epithelial cells causes molecular changes indicative of an epithelial to mesenchymal transition. Virology 391:57–63 [View Article][PubMed]
     [Google Scholar]
  22. Ishidate T., Matsumine A., Toyoshima K., Akiyama T. 2000; The APC-hDLG complex negatively regulates cell cycle progression from the G0/G1 to S phase. Oncogene 19:365–372 [View Article][PubMed]
     [Google Scholar]
  23. Jian Y., Van Tine B. A., Chien W. M., Shaw G. M., Broker T. R., Chow L. T. 1999; Concordant induction of cyclin E and p21cip1 in differentiated keratinocytes by the human papillomavirus E7 protein inhibits cellular and viral DNA synthesis. Cell Growth Differ 10:101–111[PubMed]
     [Google Scholar]
  24. Krishna Subbaiah V., Massimi P., Boon S. S., Myers M. P., Sharek L., Garcia-Mata R., Banks L. 2012; The invasive capacity of HPV transformed cells requires the hDlg-dependent enhancement of SGEF/RhoG activity. PLoS Pathog 8:e1002543 [View Article][PubMed]
     [Google Scholar]
  25. Laprise P., Viel A., Rivard N. 2004; Human homolog of disc-large is required for adherens junction assembly and differentiation of human intestinal epithelial cells. J Biol Chem 279:10157–10166 [View Article][PubMed]
     [Google Scholar]
  26. Lue R. A., Marfatia S. M., Branton D., Chishti A. H. 1994; Cloning and characterization of hdlg: the human homologue of the Drosophila discs large tumor suppressor binds to protein 4.1. Proc Natl Acad Sci U S A 91:9818–9822 [View Article][PubMed]
     [Google Scholar]
  27. Massimi P., Narayan N., Cuenda A., Banks L. 2006; Phosphorylation of the discs large tumour suppressor protein controls its membrane localisation and enhances its susceptibility to HPV E6-induced degradation. Oncogene 25:4276–4285 [View Article][PubMed]
     [Google Scholar]
  28. Matlashewski G., Schneider J., Banks L., Jones N., Murray A., Crawford L. 1987; Human papillomavirus type 16 DNA cooperates with activated ras in transforming primary cells. EMBO J 6:1741–1746[PubMed]
     [Google Scholar]
  29. McCord C., Xu J., Xu W., Qiu X., Muhanna N., Irish J., Leong I., McComb R. J., Perez-Ordonez B., Bradley G. 2014; Association of human papilloma virus with atypical and malignant oral papillary lesions. Oral Surg Oral Med Oral Pathol Oral Radiol 117:722–732 [View Article][PubMed]
     [Google Scholar]
  30. Münger K., Basile J. R., Duensing S., Eichten A., Gonzalez S. L., Grace M., Zacny V. L. 2001; Biological activities and molecular targets of the human papillomavirus E7 oncoprotein. Oncogene 20:7888–7898 [View Article][PubMed]
     [Google Scholar]
  31. Narayan N., Massimi P., Banks L. 2009; CDK phosphorylation of the discs large tumour suppressor controls its localisation and stability. J Cell Sci 122:65–74 [View Article][PubMed]
     [Google Scholar]
  32. Pei X. F., Sherman L., Sun Y. H., Schlegel R. 1998; HPV-16 E7 protein bypasses keratinocyte growth inhibition by serum and calcium. Carcinogenesis 19:1481–1486 [View Article][PubMed]
     [Google Scholar]
  33. Pim D., Banks L. 2010; Interaction of viral oncoproteins with cellular target molecules: infection with high-risk vs low-risk human papillomaviruses. APMIS 118:471–493 [CrossRef]
     [Google Scholar]
  34. Pim D., Bergant M., Boon S. S., Ganti K., Kranjec C., Massimi P., Subbaiah V. K., Thomas M., Tomaić V., Banks L. 2012; Human papillomaviruses and the specificity of PDZ domain targeting. FEBS J 279:3530–3537 [View Article][PubMed]
     [Google Scholar]
  35. Pinheiro C., Garcia E. A., Morais-Santos F., Scapulatempo-Neto C., Mafra A., Steenbergen R. D., Boccardo E., Villa L. L., Baltazar F., Longatto-Filho A. 2014; Lactate transporters and vascular factors in HPV-induced squamous cell carcinoma of the uterine cervix. BMC Cancer 14:751 [View Article][PubMed]
     [Google Scholar]
  36. Polette M., Gilles C., Nawrocki-Raby B., Lohi J., Hunziker W., Foidart J. M., Birembaut P. 2005; Membrane-type 1 matrix metalloproteinase expression is regulated by zonula occludens-1 in human breast cancer cells. Cancer Res 65:7691–7698[PubMed]
     [Google Scholar]
  37. Reuver S. M., Garner C. C. 1998; E-cadherin mediated cell adhesion recruits SAP97 into the cortical cytoskeleton. J Cell Sci 111:1071–1080[PubMed]
     [Google Scholar]
  38. Roberts S., Delury C., Marsh E. 2012; The PDZ protein discs-large (DLG): the ‘Jekyll and Hyde’ of the epithelial polarity proteins. FEBS J 279:3549–3558 [View Article][PubMed]
     [Google Scholar]
  39. Sabio G., Arthur J. S., Kuma Y., Peggie M., Carr J., Murray-Tait V., Centeno F., Goedert M., Morrice N. A., Cuenda A. 2005; p38(regulates the localisation of SAP97 in the cytoskeleton by modulating its interaction with GKAP. EMBO J 24:1134–1145 [View Article][PubMed]
     [Google Scholar]
  40. Seavey S. E., Holubar M., Saucedo L. J., Perry M. E. 1999; The E7 oncoprotein of human papillomavirus type 16 stabilizes p53 through a mechanism independent of p19(ARF). J Virol 73:7590–7598[PubMed]
     [Google Scholar]
  41. Sotelo N. S., Valiente M., Gil A., Pulido R. 2012; A functional network of the tumor suppressors APC, hDlg, and PTEN, that relies on recognition of specific PDZ-domains. J Cell Biochem 113:2661–2670 [View Article][PubMed]
     [Google Scholar]
  42. Steenbergen R. D., Parker J. N., Isern S., Snijders P. J., Walboomers J. M., Meijer C. J., Broker T. R., Chow L. T. 1998; Viral E6-E7 transcription in the basal layer of organotypic cultures without apparent p21cip1 protein precedes immortalization of human papillomavirus type 16- and 18-transfected human keratinocytes. J Virol 72:749–757[PubMed]
     [Google Scholar]
  43. Thomas J. T., Laimins L. A. 1998; Human papillomavirus oncoproteins E6 and E7 independently abrogate the mitotic spindle checkpoint. J Virol 72:1131–1137[PubMed]
     [Google Scholar]
  44. Thomas J. T., Oh S. T., Terhune S. S., Laimins L. A. 2001; Cellular changes induced by low-risk human papillomavirus type 11 in keratinocytes that stably maintain viral episomes. J Virol 75:7564–7571 [View Article][PubMed]
     [Google Scholar]
  45. Thomas M., Dasgupta J., Zhang Y., Chen X., Banks L. 2008; Analysis of specificity determinants in the interactions of different HPV E6 proteins with their PDZ domain-containing substrates. Virology 376:371–378 [View Article][PubMed]
     [Google Scholar]
  46. Valiente M., Andrés-Pons A., Gomar B., Torres J., Gil A., Tapparel C., Antonarakis S. E., Pulido R. 2005; Binding of PTEN to specific PDZ domains contributes to PTEN protein stability and phosphorylation by microtubule-associated serine/threonine kinases. J Biol Chem 280:28936–28943 [View Article][PubMed]
     [Google Scholar]
  47. Wang H. K., Duffy A. A., Broker T. R., Chow L. T. 2009; Robust production and passaging of infectious HPV in squamous epithelium of primary human keratinocytes. Genes Dev 23:181–194 [View Article][PubMed]
     [Google Scholar]
  48. Watson R. A., Rollason T. P., Reynolds G. M., Murray P. G., Banks L., Roberts S. 2002; Changes in expression of the human homologue of the Drosophila discs large tumour suppressor protein in high-grade premalignant cervical neoplasias. Carcinogenesis 23:1791–1796 [View Article][PubMed]
     [Google Scholar]
  49. Zehbe I., Richard C., DeCarlo C. A., Shai A., Lambert P. F., Lichtig H., Tommasino M., Sherman L. 2009; Human papillomavirus 16 E6 variants differ in their dysregulation of human keratinocyte differentiation and apoptosis. Virology 383:69–77 [View Article][PubMed]
     [Google Scholar]
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Disc large 1 expression is altered by human papillomavirus E6/E7 proteins in organotypic cultures of human keratinocytes
J. Gen. Virol. 97, 453 (2016); https://doi.org/10.1099/jgv.0.000364
/content/journal/jgv/10.1099/jgv.0.000364
/content/journal/jgv/10.1099/jgv.0.000364
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