Skip to main content
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1993 Mar 11;21(5):1103–1109. doi: 10.1093/nar/21.5.1103

Ultraviolet A (320-380 nm) radiation causes an alteration in the binding of a specific protein/protein complex to a short region of the promoter of the human heme oxygenase 1 gene.

A L Nascimento 1, P Luscher 1, R M Tyrrell 1
PMCID: PMC309269  PMID: 7840819

Abstract

Ultraviolet A (320-380 nm) radiation strongly stimulates expression of the human heme oxygenase 1 gene as a consequence of an enhancement in transcription rate (1). We have used a 147 bp fragment of the promoter of this gene as a probe for DNA binding activity in nuclear extracts prepared from untreated and UVA treated populations of cultured human skin fibroblasts. Analysis using gel electrophoresis mobility shift assays clearly demonstrates the appearance of a strong binding activity unique to UVA-treated extracts that is formed in the absence of de novo protein synthesis. Footprint analysis defines a binding region from -41 to -50 bp that partially overlaps with a region known to constitutively bind upstream stimulatory factor (USF). Further analysis using synthetic oligonucleotides and gel retardation has confirmed that the crucial sequence for binding the protein present in both control and UVA-treated extracts lies within a 26 bp sequence that includes the core USF binding site. UVA radiation appears to lead to a modification of the USF complex (or closely related proteins(s)) to give a structurally modified protein/protein complex which protects only the upstream half of a region originally defined by foot-printing of the promoter with USF.

Full text

PDF
1103

Images in this article

Selected References

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

  1. Applegate L. A., Luscher P., Tyrrell R. M. Induction of heme oxygenase: a general response to oxidant stress in cultured mammalian cells. Cancer Res. 1991 Feb 1;51(3):974–978. [PubMed] [Google Scholar]
  2. Ben-Hattar J., Beard P., Jiricny J. Cytosine methylation in CTF and Sp1 recognition sites of an HSV tk promoter: effects on transcription in vivo and on factor binding in vitro. Nucleic Acids Res. 1989 Dec 25;17(24):10179–10190. doi: 10.1093/nar/17.24.10179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Benton W. D., Davis R. W. Screening lambdagt recombinant clones by hybridization to single plaques in situ. Science. 1977 Apr 8;196(4286):180–182. doi: 10.1126/science.322279. [DOI] [PubMed] [Google Scholar]
  4. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  5. Devary Y., Gottlieb R. A., Lau L. F., Karin M. Rapid and preferential activation of the c-jun gene during the mammalian UV response. Mol Cell Biol. 1991 May;11(5):2804–2811. doi: 10.1128/mcb.11.5.2804. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dignam J. D., Lebovitz R. M., Roeder R. G. Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucleic Acids Res. 1983 Mar 11;11(5):1475–1489. doi: 10.1093/nar/11.5.1475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Keyse S. M., Applegate L. A., Tromvoukis Y., Tyrrell R. M. Oxidant stress leads to transcriptional activation of the human heme oxygenase gene in cultured skin fibroblasts. Mol Cell Biol. 1990 Sep;10(9):4967–4969. doi: 10.1128/mcb.10.9.4967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Keyse S. M., Tyrrell R. M. Heme oxygenase is the major 32-kDa stress protein induced in human skin fibroblasts by UVA radiation, hydrogen peroxide, and sodium arsenite. Proc Natl Acad Sci U S A. 1989 Jan;86(1):99–103. doi: 10.1073/pnas.86.1.99. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Martin M. L., Kirkpatrick H. Nursing assessment of the aggressive elderly. Perspectives. 1987 Fall;11(3):8–10. [PubMed] [Google Scholar]
  10. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  11. Mihm S., Ennen J., Pessara U., Kurth R., Dröge W. Inhibition of HIV-1 replication and NF-kappa B activity by cysteine and cysteine derivatives. AIDS. 1991 May;5(5):497–503. doi: 10.1097/00002030-199105000-00004. [DOI] [PubMed] [Google Scholar]
  12. Offord E. A., Beard P. A member of the activator protein 1 family found in keratinocytes but not in fibroblasts required for transcription from a human papillomavirus type 18 promoter. J Virol. 1990 Oct;64(10):4792–4798. doi: 10.1128/jvi.64.10.4792-4798.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Riccio A., Pedone P. V., Lund L. R., Olesen T., Olsen H. S., Andreasen P. A. Transforming growth factor beta 1-responsive element: closely associated binding sites for USF and CCAAT-binding transcription factor-nuclear factor I in the type 1 plasminogen activator inhibitor gene. Mol Cell Biol. 1992 Apr;12(4):1846–1855. doi: 10.1128/mcb.12.4.1846. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Sato M., Ishizawa S., Yoshida T., Shibahara S. Interaction of upstream stimulatory factor with the human heme oxygenase gene promoter. Eur J Biochem. 1990 Mar 10;188(2):231–237. doi: 10.1111/j.1432-1033.1990.tb15394.x. [DOI] [PubMed] [Google Scholar]
  15. Sawadogo M., Sentenac A. RNA polymerase B (II) and general transcription factors. Annu Rev Biochem. 1990;59:711–754. doi: 10.1146/annurev.bi.59.070190.003431. [DOI] [PubMed] [Google Scholar]
  16. Schreck R., Rieber P., Baeuerle P. A. Reactive oxygen intermediates as apparently widely used messengers in the activation of the NF-kappa B transcription factor and HIV-1. EMBO J. 1991 Aug;10(8):2247–2258. doi: 10.1002/j.1460-2075.1991.tb07761.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Shibahara S., Sato M., Muller R. M., Yoshida T. Structural organization of the human heme oxygenase gene and the function of its promoter. Eur J Biochem. 1989 Feb 15;179(3):557–563. doi: 10.1111/j.1432-1033.1989.tb14583.x. [DOI] [PubMed] [Google Scholar]
  18. Sorger P. K. Heat shock factor and the heat shock response. Cell. 1991 May 3;65(3):363–366. doi: 10.1016/0092-8674(91)90452-5. [DOI] [PubMed] [Google Scholar]
  19. Tyrrell R. M., Pidoux M. Quantitative differences in host cell reactivation of ultraviolet-damaged virus in human skin fibroblasts and epidermal keratinocytes cultured from the same foreskin biopsy. Cancer Res. 1986 Jun;46(6):2665–2669. [PubMed] [Google Scholar]
  20. Zabel U., Schreck R., Baeuerle P. A. DNA binding of purified transcription factor NF-kappa B. Affinity, specificity, Zn2+ dependence, and differential half-site recognition. J Biol Chem. 1991 Jan 5;266(1):252–260. [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES