Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1993 Jun 1;121(5):1109–1120. doi: 10.1083/jcb.121.5.1109

Skin-specific expression of a truncated E1a oncoprotein binding to p105- Rb leads to abnormal hair follicle maturation without increased epidermal proliferation

PMCID: PMC2119685  PMID: 7684738

Abstract

In cultured cells, mutants of the Adenovirus E1a oncoprotein which bind to a reduced set of cellular proteins, including p105-Rb, p107, and p60- cyclin A, are transformation defective but can still interfere with exogenous growth inhibitory and differentiating signals, such as those triggered by TGF-beta. We have tested the ability of one such mutant, NTdl646, to interfere with keratinocyte growth and differentiation in vivo, in the skin of transgenic mice. Keratinocyte-specific expression of the transgene was achieved by using a keratin 5 promoter. Two independent lines of transgenic mice were obtained which expressed E1a specifically in their skin and exhibited an aberrant hair coat phenotype with striking regional variations. Affected hair shafts were short and crooked and hair follicles exhibited a dystrophic or absent inner root sheath. Interfollicular epidermis was normal, but its hyperplastic response to acute treatment with TPA (12-O- tetradecanoylphorbol-13-acetate) was significantly reduced. Primary keratinocytes derived from these animals were partially resistant to the effects of TPA and TGF-beta. The rate of spontaneous or chemically induced skin tumors in the transgenic mice was not increased. Thus, expression of a transgene which interferes with known negative growth regulatory proteins causes profound disturbances of keratinocyte maturation into a highly organized structure such as the hair follicle but does not lead to increased and/or neoplastic proliferation.

Full Text

The Full Text of this article is available as a PDF (7.0 MB).

Selected References

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

  1. Argyris T. S. Tumor promotion by abrasion induced epidermal hyperplasia in the skin of mice. J Invest Dermatol. 1980 Oct;75(4):360–362. doi: 10.1111/1523-1747.ep12531153. [DOI] [PubMed] [Google Scholar]
  2. BERENBLUM I., HARAN-GHERA N., TRAININ N. An experimental analysis of the hair cycle effect in mouse skin carcinogenesis. Br J Cancer. 1958 Sep;12(3):402–413. doi: 10.1038/bjc.1958.48. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bailleul B., Surani M. A., White S., Barton S. C., Brown K., Blessing M., Jorcano J., Balmain A. Skin hyperkeratosis and papilloma formation in transgenic mice expressing a ras oncogene from a suprabasal keratin promoter. Cell. 1990 Aug 24;62(4):697–708. doi: 10.1016/0092-8674(90)90115-u. [DOI] [PubMed] [Google Scholar]
  4. Balmain A., Ramsden M., Bowden G. T., Smith J. Activation of the mouse cellular Harvey-ras gene in chemically induced benign skin papillomas. Nature. 1984 Feb 16;307(5952):658–660. doi: 10.1038/307658a0. [DOI] [PubMed] [Google Scholar]
  5. Barrandon Y., Morgan J. R., Mulligan R. C., Green H. Restoration of growth potential in paraclones of human keratinocytes by a viral oncogene. Proc Natl Acad Sci U S A. 1989 Jun;86(11):4102–4106. doi: 10.1073/pnas.86.11.4102. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bhisey R. A., Iyengar B., Sirsat S. M. Effect of the active tumor promoter, 12-O-tetradecanoylphorbol-13-acetate on hair follicular growth and development of hair anlage tumors in the mouse skin: a comparison with human adnexal lesions. J Cancer Res Clin Oncol. 1982;102(3):245–252. doi: 10.1007/BF00411344. [DOI] [PubMed] [Google Scholar]
  7. Billingham R. E., Silvers W. K. Studies on the conservation of epidermal specificies of skin and certain mucosas in adult mammals. J Exp Med. 1967 Mar 1;125(3):429–446. doi: 10.1084/jem.125.3.429. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Boulanger P. A., Blair G. E. Expression and interactions of human adenovirus oncoproteins. Biochem J. 1991 Apr 15;275(Pt 2):281–299. doi: 10.1042/bj2750281. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Cao L., Faha B., Dembski M., Tsai L. H., Harlow E., Dyson N. Independent binding of the retinoblastoma protein and p107 to the transcription factor E2F. Nature. 1992 Jan 9;355(6356):176–179. doi: 10.1038/355176a0. [DOI] [PubMed] [Google Scholar]
  10. Chellappan S. P., Hiebert S., Mudryj M., Horowitz J. M., Nevins J. R. The E2F transcription factor is a cellular target for the RB protein. Cell. 1991 Jun 14;65(6):1053–1061. doi: 10.1016/0092-8674(91)90557-f. [DOI] [PubMed] [Google Scholar]
  11. Chiu R., Boyle W. J., Meek J., Smeal T., Hunter T., Karin M. The c-Fos protein interacts with c-Jun/AP-1 to stimulate transcription of AP-1 responsive genes. Cell. 1988 Aug 12;54(4):541–552. doi: 10.1016/0092-8674(88)90076-1. [DOI] [PubMed] [Google Scholar]
  12. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  13. Clemmensen O. J., Hainau B., Hansted B. The ultrastructure of the transition zone between specialized cells ("Flügelzellen") of Huxley's layer of the inner root sheath and cells of the outer root sheath of the human hair follicle. Am J Dermatopathol. 1991 Jun;13(3):264–270. doi: 10.1097/00000372-199106000-00008. [DOI] [PubMed] [Google Scholar]
  14. Cotsarelis G., Sun T. T., Lavker R. M. Label-retaining cells reside in the bulge area of pilosebaceous unit: implications for follicular stem cells, hair cycle, and skin carcinogenesis. Cell. 1990 Jun 29;61(7):1329–1337. doi: 10.1016/0092-8674(90)90696-c. [DOI] [PubMed] [Google Scholar]
  15. Coulomb B., Lebreton C., Dubertret L. Influence of human dermal fibroblasts on epidermalization. J Invest Dermatol. 1989 Jan;92(1):122–125. doi: 10.1111/1523-1747.ep13071335. [DOI] [PubMed] [Google Scholar]
  16. Donehower L. A., Harvey M., Slagle B. L., McArthur M. J., Montgomery C. A., Jr, Butel J. S., Bradley A. Mice deficient for p53 are developmentally normal but susceptible to spontaneous tumours. Nature. 1992 Mar 19;356(6366):215–221. doi: 10.1038/356215a0. [DOI] [PubMed] [Google Scholar]
  17. Ewen M. E., Faha B., Harlow E., Livingston D. M. Interaction of p107 with cyclin A independent of complex formation with viral oncoproteins. Science. 1992 Jan 3;255(5040):85–87. doi: 10.1126/science.1532457. [DOI] [PubMed] [Google Scholar]
  18. Filvaroff E., Stern D. F., Dotto G. P. Tyrosine phosphorylation is an early and specific event involved in primary keratinocyte differentiation. Mol Cell Biol. 1990 Mar;10(3):1164–1173. doi: 10.1128/mcb.10.3.1164. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Fisher C., Byers M. R., Iadarola M. J., Powers E. A. Patterns of epithelial expression of Fos protein suggest important role in the transition from viable to cornified cell during keratinization. Development. 1991 Feb;111(2):253–258. doi: 10.1242/dev.111.2.253. [DOI] [PubMed] [Google Scholar]
  20. Florin-Christensen M., Missero C., Dotto G. P., Florin-Christensen J. The E1a gene prevents inhibition of keratinocyte proliferation by dexamethasone. Exp Cell Res. 1992 Nov;203(1):285–288. doi: 10.1016/0014-4827(92)90067-i. [DOI] [PubMed] [Google Scholar]
  21. Fuchs E. Epidermal differentiation: the bare essentials. J Cell Biol. 1990 Dec;111(6 Pt 2):2807–2814. doi: 10.1083/jcb.111.6.2807. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Fuchs E., Green H. Changes in keratin gene expression during terminal differentiation of the keratinocyte. Cell. 1980 Apr;19(4):1033–1042. doi: 10.1016/0092-8674(80)90094-x. [DOI] [PubMed] [Google Scholar]
  23. Greenhalgh D. A., Yuspa S. H. Malignant conversion of murine squamous papilloma cell lines by transfection with the fos oncogene. Mol Carcinog. 1988;1(2):134–143. doi: 10.1002/mc.2940010209. [DOI] [PubMed] [Google Scholar]
  24. Heid H. W., Moll I., Franke W. W. Patterns of expression of trichocytic and epithelial cytokeratins in mammalian tissues. I. Human and bovine hair follicles. Differentiation. 1988;37(2):137–157. doi: 10.1111/j.1432-0436.1988.tb00805.x. [DOI] [PubMed] [Google Scholar]
  25. Heine U., Munoz E. F., Flanders K. C., Ellingsworth L. R., Lam H. Y., Thompson N. L., Roberts A. B., Sporn M. B. Role of transforming growth factor-beta in the development of the mouse embryo. J Cell Biol. 1987 Dec;105(6 Pt 2):2861–2876. doi: 10.1083/jcb.105.6.2861. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Hennings H., Devor D., Wenk M. L., Slaga T. J., Former B., Colburn N. H., Bowden G. T., Elgjo K., Yuspa S. H. Comparison of two-stage epidermal carcinogenesis initiated by 7,12-dimethylbenz(a)anthracene or N-methyl-N'-nitro-N-nitrosoguanidine in newborn and adult SENCAR and BALB/c mice. Cancer Res. 1981 Mar;41(3):773–779. [PubMed] [Google Scholar]
  27. Hennings H., Michael D., Cheng C., Steinert P., Holbrook K., Yuspa S. H. Calcium regulation of growth and differentiation of mouse epidermal cells in culture. Cell. 1980 Jan;19(1):245–254. doi: 10.1016/0092-8674(80)90406-7. [DOI] [PubMed] [Google Scholar]
  28. Hirai Y., Takebe K., Takashina M., Kobayashi S., Takeichi M. Epimorphin: a mesenchymal protein essential for epithelial morphogenesis. Cell. 1992 May 1;69(3):471–481. doi: 10.1016/0092-8674(92)90448-l. [DOI] [PubMed] [Google Scholar]
  29. Inaba M., Anthony J., McKinstry C. Histologic study of the regeneration of axillary hair after removal with subcutaneous tissue shaver. J Invest Dermatol. 1979 May;72(5):224–231. doi: 10.1111/1523-1747.ep12530773. [DOI] [PubMed] [Google Scholar]
  30. Jacks T., Fazeli A., Schmitt E. M., Bronson R. T., Goodell M. A., Weinberg R. A. Effects of an Rb mutation in the mouse. Nature. 1992 Sep 24;359(6393):295–300. doi: 10.1038/359295a0. [DOI] [PubMed] [Google Scholar]
  31. Koike K., Hinrichs S. H., Isselbacher K. J., Jay G. Transgenic mouse model for human gastric carcinoma. Proc Natl Acad Sci U S A. 1989 Jul;86(14):5615–5619. doi: 10.1073/pnas.86.14.5615. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  33. Lee E. Y., Chang C. Y., Hu N., Wang Y. C., Lai C. C., Herrup K., Lee W. H., Bradley A. Mice deficient for Rb are nonviable and show defects in neurogenesis and haematopoiesis. Nature. 1992 Sep 24;359(6393):288–294. doi: 10.1038/359288a0. [DOI] [PubMed] [Google Scholar]
  34. Lenoir M. C., Bernard B. A., Pautrat G., Darmon M., Shroot B. Outer root sheath cells of human hair follicle are able to regenerate a fully differentiated epidermis in vitro. Dev Biol. 1988 Dec;130(2):610–620. doi: 10.1016/0012-1606(88)90356-9. [DOI] [PubMed] [Google Scholar]
  35. Lersch R., Stellmach V., Stocks C., Giudice G., Fuchs E. Isolation, sequence, and expression of a human keratin K5 gene: transcriptional regulation of keratins and insights into pairwise control. Mol Cell Biol. 1989 Sep;9(9):3685–3697. doi: 10.1128/mcb.9.9.3685. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Limat A., Breitkreutz D., Hunziker T., Boillat C., Wiesmann U., Klein E., Noser F., Fusenig N. E. Restoration of the epidermal phenotype by follicular outer root sheath cells in recombinant culture with dermal fibroblasts. Exp Cell Res. 1991 Jun;194(2):218–227. doi: 10.1016/0014-4827(91)90357-z. [DOI] [PubMed] [Google Scholar]
  37. Mintz B., Bradl M. Mosaic expression of a tyrosinase fusion gene in albino mice yields a heritable striped coat color pattern in transgenic homozygotes. Proc Natl Acad Sci U S A. 1991 Nov 1;88(21):9643–9647. doi: 10.1073/pnas.88.21.9643. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Missero C., Filvaroff E., Dotto G. P. Induction of transforming growth factor beta 1 resistance by the E1A oncogene requires binding to a specific set of cellular proteins. Proc Natl Acad Sci U S A. 1991 Apr 15;88(8):3489–3493. doi: 10.1073/pnas.88.8.3489. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Missero C., Ramon y Cajal S., Dotto G. P. Escape from transforming growth factor beta control and oncogene cooperation in skin tumor development. Proc Natl Acad Sci U S A. 1991 Nov 1;88(21):9613–9617. doi: 10.1073/pnas.88.21.9613. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Moll R., Franke W. W., Schiller D. L., Geiger B., Krepler R. The catalog of human cytokeratins: patterns of expression in normal epithelia, tumors and cultured cells. Cell. 1982 Nov;31(1):11–24. doi: 10.1016/0092-8674(82)90400-7. [DOI] [PubMed] [Google Scholar]
  41. Moran E., Mathews M. B. Multiple functional domains in the adenovirus E1A gene. Cell. 1987 Jan 30;48(2):177–178. doi: 10.1016/0092-8674(87)90418-1. [DOI] [PubMed] [Google Scholar]
  42. Mudryj M., Devoto S. H., Hiebert S. W., Hunter T., Pines J., Nevins J. R. Cell cycle regulation of the E2F transcription factor involves an interaction with cyclin A. Cell. 1991 Jun 28;65(7):1243–1253. doi: 10.1016/0092-8674(91)90019-u. [DOI] [PubMed] [Google Scholar]
  43. Nevins J. R. E2F: a link between the Rb tumor suppressor protein and viral oncoproteins. Science. 1992 Oct 16;258(5081):424–429. doi: 10.1126/science.1411535. [DOI] [PubMed] [Google Scholar]
  44. Oliver R. F. Whisker growth after removal of the dermal papilla and lengths of follicle in the hooded rat. J Embryol Exp Morphol. 1966 Jun;15(3):331–347. [PubMed] [Google Scholar]
  45. Pelton R. W., Saxena B., Jones M., Moses H. L., Gold L. I. Immunohistochemical localization of TGF beta 1, TGF beta 2, and TGF beta 3 in the mouse embryo: expression patterns suggest multiple roles during embryonic development. J Cell Biol. 1991 Nov;115(4):1091–1105. doi: 10.1083/jcb.115.4.1091. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Pietenpol J. A., Stein R. W., Moran E., Yaciuk P., Schlegel R., Lyons R. M., Pittelkow M. R., Münger K., Howley P. M., Moses H. L. TGF-beta 1 inhibition of c-myc transcription and growth in keratinocytes is abrogated by viral transforming proteins with pRB binding domains. Cell. 1990 Jun 1;61(5):777–785. doi: 10.1016/0092-8674(90)90188-k. [DOI] [PubMed] [Google Scholar]
  47. Powell B. C., Rogers G. E. Cyclic hair-loss and regrowth in transgenic mice overexpressing an intermediate filament gene. EMBO J. 1990 May;9(5):1485–1493. doi: 10.1002/j.1460-2075.1990.tb08266.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Reiners J. J., Jr, Slaga T. J. Effects of tumor promoters on the rate and commitment to terminal differentiation of subpopulations of murine keratinocytes. Cell. 1983 Jan;32(1):247–255. doi: 10.1016/0092-8674(83)90515-9. [DOI] [PubMed] [Google Scholar]
  49. Rogers G. E., Fietz M. J., Fratini A. Trichohyalin and matrix proteins. Ann N Y Acad Sci. 1991 Dec 26;642:64–81. [PubMed] [Google Scholar]
  50. STRAILE W. E. Possible functions of the external root sheath during growth of the hair follicle. J Exp Zool. 1962 Aug;150:207–223. doi: 10.1002/jez.1401500305. [DOI] [PubMed] [Google Scholar]
  51. Saiag P., Coulomb B., Lebreton C., Bell E., Dubertret L. Psoriatic fibroblasts induce hyperproliferation of normal keratinocytes in a skin equivalent model in vitro. Science. 1985 Nov 8;230(4726):669–672. doi: 10.1126/science.2413549. [DOI] [PubMed] [Google Scholar]
  52. Sassone-Corsi P., Lamph W. W., Kamps M., Verma I. M. fos-associated cellular p39 is related to nuclear transcription factor AP-1. Cell. 1988 Aug 12;54(4):553–560. doi: 10.1016/0092-8674(88)90077-3. [DOI] [PubMed] [Google Scholar]
  53. Schweizer J., Marks F. Induction of the formation of new hair follicles in mouse tail epidermis by the tumor promoter 12-O-tetradecanoylphorbol-13-acetate. Cancer Res. 1977 Nov;37(11):4195–4201. [PubMed] [Google Scholar]
  54. Southern P. J., Berg P. Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter. J Mol Appl Genet. 1982;1(4):327–341. [PubMed] [Google Scholar]
  55. Stanbridge E. J. Human tumor suppressor genes. Annu Rev Genet. 1990;24:615–657. doi: 10.1146/annurev.ge.24.120190.003151. [DOI] [PubMed] [Google Scholar]
  56. Steinert P., Yuspa S. H. Biochemical evidence for keratinization by mouse epidermal cells in culture. Science. 1978 Jun 30;200(4349):1491–1493. doi: 10.1126/science.566466. [DOI] [PubMed] [Google Scholar]
  57. Tso J. Y., Sun X. H., Kao T. H., Reece K. S., Wu R. Isolation and characterization of rat and human glyceraldehyde-3-phosphate dehydrogenase cDNAs: genomic complexity and molecular evolution of the gene. Nucleic Acids Res. 1985 Apr 11;13(7):2485–2502. doi: 10.1093/nar/13.7.2485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. Vassar R., Fuchs E. Transgenic mice provide new insights into the role of TGF-alpha during epidermal development and differentiation. Genes Dev. 1991 May;5(5):714–727. doi: 10.1101/gad.5.5.714. [DOI] [PubMed] [Google Scholar]
  59. Whyte P., Williamson N. M., Harlow E. Cellular targets for transformation by the adenovirus E1A proteins. Cell. 1989 Jan 13;56(1):67–75. doi: 10.1016/0092-8674(89)90984-7. [DOI] [PubMed] [Google Scholar]
  60. Wilson J. B., Weinberg W., Johnson R., Yuspa S., Levine A. J. Expression of the BNLF-1 oncogene of Epstein-Barr virus in the skin of transgenic mice induces hyperplasia and aberrant expression of keratin 6. Cell. 1990 Jun 29;61(7):1315–1327. doi: 10.1016/0092-8674(90)90695-b. [DOI] [PubMed] [Google Scholar]
  61. Yamasaki H. Gap junctional intercellular communication and carcinogenesis. Carcinogenesis. 1990 Jul;11(7):1051–1058. doi: 10.1093/carcin/11.7.1051. [DOI] [PubMed] [Google Scholar]
  62. Yuspa S. H., Ben T., Hennings H., Lichti U. Divergent responses in epidermal basal cells exposed to the tumor promoter 12-O-tetradecanoylphorbol-13-acetate. Cancer Res. 1982 Jun;42(6):2344–2349. [PubMed] [Google Scholar]
  63. Yuspa S. H., Hawley-Nelson P., Koehler B., Stanley J. R. A survey of transformation markers in differentiating epidermal cell lines in culture. Cancer Res. 1980 Dec;40(12):4694–4703. [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES