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. 1990 Aug 11;18(15):4401–4407. doi: 10.1093/nar/18.15.4401

Characterization of the human spr2 promoter: induction after UV irradiation or TPA treatment and regulation during differentiation of cultured primary keratinocytes.

S Gibbs 1, F Lohman 1, W Teubel 1, P van de Putte 1, C Backendorf 1
PMCID: PMC331257  PMID: 2388825

Abstract

We have isolated genomic clones from several members of the UV and TPA inducible human spr2 gene-family in order to analyse the regulation of these genes at a molecular level. From one of these members, the spr2-1 gene, we have identified and sequenced the regulatory region. By using CAT fusion plasmids and a liposome mediated transfection procedure we show that the isolated promoter region contains all the cis-elements necessary for induced expression after UV irradiation or phorbolester treatment of cultured human keratinocytes. Additionally the spr2-1 promoter is shown to be regulated aswell during the normal process of keratinocyte differentiation. This makes the spr2-1 promoter sequence an ideal tool to study the molecular mechanisms by which environmental agents such as UV radiation and chemical tumor promoters interfere with normal gene expression during cell proliferation and differentiation.

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Selected References

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

  1. Angel P., Baumann I., Stein B., Delius H., Rahmsdorf H. J., Herrlich P. 12-O-tetradecanoyl-phorbol-13-acetate induction of the human collagenase gene is mediated by an inducible enhancer element located in the 5'-flanking region. Mol Cell Biol. 1987 Jun;7(6):2256–2266. doi: 10.1128/mcb.7.6.2256. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Angel P., Imagawa M., Chiu R., Stein B., Imbra R. J., Rahmsdorf H. J., Jonat C., Herrlich P., Karin M. Phorbol ester-inducible genes contain a common cis element recognized by a TPA-modulated trans-acting factor. Cell. 1987 Jun 19;49(6):729–739. doi: 10.1016/0092-8674(87)90611-8. [DOI] [PubMed] [Google Scholar]
  3. Belt P. B., Groeneveld H., Teubel W. J., van de Putte P., Backendorf C. Construction and properties of an Epstein-Barr-virus-derived cDNA expression vector for human cells. Gene. 1989 Dec 14;84(2):407–417. doi: 10.1016/0378-1119(89)90515-5. [DOI] [PubMed] [Google Scholar]
  4. Büscher M., Rahmsdorf H. J., Litfin M., Karin M., Herrlich P. Activation of the c-fos gene by UV and phorbol ester: different signal transduction pathways converge to the same enhancer element. Oncogene. 1988 Sep;3(3):301–311. [PubMed] [Google Scholar]
  5. Curran T., Franza B. R., Jr Fos and Jun: the AP-1 connection. Cell. 1988 Nov 4;55(3):395–397. doi: 10.1016/0092-8674(88)90024-4. [DOI] [PubMed] [Google Scholar]
  6. Dynan W. S. Modularity in promoters and enhancers. Cell. 1989 Jul 14;58(1):1–4. doi: 10.1016/0092-8674(89)90393-0. [DOI] [PubMed] [Google Scholar]
  7. Elwood J. M., Whitehead S. M., Gallagher R. P. Epidemiology of human malignant skin tumors with special reference to natural and artificial ultraviolet radiation exposures. Carcinog Compr Surv. 1989;11:55–84. [PubMed] [Google Scholar]
  8. Felgner P. L., Gadek T. R., Holm M., Roman R., Chan H. W., Wenz M., Northrop J. P., Ringold G. M., Danielsen M. Lipofection: a highly efficient, lipid-mediated DNA-transfection procedure. Proc Natl Acad Sci U S A. 1987 Nov;84(21):7413–7417. doi: 10.1073/pnas.84.21.7413. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gorman C. M., Moffat L. F., Howard B. H. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol. 1982 Sep;2(9):1044–1051. doi: 10.1128/mcb.2.9.1044. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Graham F. L., van der Eb A. J. Transformation of rat cells by DNA of human adenovirus 5. Virology. 1973 Aug;54(2):536–539. doi: 10.1016/0042-6822(73)90163-3. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Kartasova T., Cornelissen B. J., Belt P., van de Putte P. Effects of UV, 4-NQO and TPA on gene expression in cultured human epidermal keratinocytes. Nucleic Acids Res. 1987 Aug 11;15(15):5945–5962. doi: 10.1093/nar/15.15.5945. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kartasova T., van Muijen G. N., van Pelt-Heerschap H., van de Putte P. Novel protein in human epidermal keratinocytes: regulation of expression during differentiation. Mol Cell Biol. 1988 May;8(5):2204–2210. doi: 10.1128/mcb.8.5.2204. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kartasova T., van de Putte P. Isolation, characterization, and UV-stimulated expression of two families of genes encoding polypeptides of related structure in human epidermal keratinocytes. Mol Cell Biol. 1988 May;8(5):2195–2203. doi: 10.1128/mcb.8.5.2195. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Klein-Szanto A. J. Pathology of human and experimental skin tumors. Carcinog Compr Surv. 1989;11:19–53. [PubMed] [Google Scholar]
  16. Lee W., Haslinger A., Karin M., Tjian R. Activation of transcription by two factors that bind promoter and enhancer sequences of the human metallothionein gene and SV40. Nature. 1987 Jan 22;325(6102):368–372. doi: 10.1038/325368a0. [DOI] [PubMed] [Google Scholar]
  17. Marsh J. L., Erfle M., Wykes E. J. The pIC plasmid and phage vectors with versatile cloning sites for recombinant selection by insertional inactivation. Gene. 1984 Dec;32(3):481–485. doi: 10.1016/0378-1119(84)90022-2. [DOI] [PubMed] [Google Scholar]
  18. Messing J., Crea R., Seeburg P. H. A system for shotgun DNA sequencing. Nucleic Acids Res. 1981 Jan 24;9(2):309–321. doi: 10.1093/nar/9.2.309. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Mount S. M. A catalogue of splice junction sequences. Nucleic Acids Res. 1982 Jan 22;10(2):459–472. doi: 10.1093/nar/10.2.459. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Ponec M., Kempenaar J. A., De Kloet E. R. Corticoids and cultured human epidermal keratinocytes: specific intracellular binding and clinical efficacy. J Invest Dermatol. 1981 Mar;76(3):211–214. doi: 10.1111/1523-1747.ep12525761. [DOI] [PubMed] [Google Scholar]
  21. Quintanilla M., Brown K., Ramsden M., Balmain A. Carcinogen-specific mutation and amplification of Ha-ras during mouse skin carcinogenesis. Nature. 1986 Jul 3;322(6074):78–80. doi: 10.1038/322078a0. [DOI] [PubMed] [Google Scholar]
  22. Rheinwald J. G., Green H. Serial cultivation of strains of human epidermal keratinocytes: the formation of keratinizing colonies from single cells. Cell. 1975 Nov;6(3):331–343. doi: 10.1016/s0092-8674(75)80001-8. [DOI] [PubMed] [Google Scholar]
  23. Siegenthaler G., Saurat J. H., Ponec M. Terminal differentiation in cultured human keratinocytes is associated with increased levels of cellular retinoic acid-binding protein. Exp Cell Res. 1988 Sep;178(1):114–126. doi: 10.1016/0014-4827(88)90383-7. [DOI] [PubMed] [Google Scholar]
  24. Staberg B., Wulf H. C., Klemp P., Poulsen T., Brodthagen H. The carcinogenic effect of UVA irradiation. J Invest Dermatol. 1983 Dec;81(6):517–519. doi: 10.1111/1523-1747.ep12522855. [DOI] [PubMed] [Google Scholar]
  25. Stein B., Rahmsdorf H. J., Steffen A., Litfin M., Herrlich P. UV-induced DNA damage is an intermediate step in UV-induced expression of human immunodeficiency virus type 1, collagenase, c-fos, and metallothionein. Mol Cell Biol. 1989 Nov;9(11):5169–5181. doi: 10.1128/mcb.9.11.5169. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Strickland P. T. Photocarcinogenesis by near-ultraviolet (UVA) radiation in Sencar mice. J Invest Dermatol. 1986 Aug;87(2):272–275. doi: 10.1111/1523-1747.ep12696669. [DOI] [PubMed] [Google Scholar]
  27. Suarez H. G., Daya-Grosjean L., Schlaifer D., Nardeux P., Renault G., Bos J. L., Sarasin A. Activated oncogenes in human skin tumors from a repair-deficient syndrome, xeroderma pigmentosum. Cancer Res. 1989 Mar 1;49(5):1223–1228. [PubMed] [Google Scholar]
  28. Takebe H., Nii S., Ishii M. I., Utsumi H. Comparative studies of host-cell reactivation, colony forming ability and excision repair after UV irradiation of xeroderma pigmentosum, normal human and some other mammalian cells. Mutat Res. 1974 Dec;25(3):383–390. doi: 10.1016/0027-5107(74)90067-0. [DOI] [PubMed] [Google Scholar]
  29. Vaheri A., Pagano J. S. Infectious poliovirus RNA: a sensitive method of assay. Virology. 1965 Nov;27(3):434–436. doi: 10.1016/0042-6822(65)90126-1. [DOI] [PubMed] [Google Scholar]
  30. Watt F. M., Green H. Stratification and terminal differentiation of cultured epidermal cells. Nature. 1982 Feb 4;295(5848):434–436. doi: 10.1038/295434a0. [DOI] [PubMed] [Google Scholar]

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