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. 1994 Jan;93(1):56–62. doi: 10.1172/JCI116984

Analysis of tumor necrosis factor promoter responses to ultraviolet light.

F Bazzoni 1, V Kruys 1, A Shakhov 1, C V Jongeneel 1, B Beutler 1
PMCID: PMC293726  PMID: 8282822

Abstract

Ultraviolet (UV) light induces the biosynthesis of chloramphenicol acetyltransferase (CAT) in the skin of mice bearing the CATTNF reporter transgene. Moreover, nuclear run-on assays indicate that UV light induces transcription of the TNF gene in RAW 264.7 macrophages. These observations suggest that the TNF gene (and/or its mRNA product) responds to signals elicited by UV light. To identify transcriptional UV response elements within the TNF promoter, and to determine whether a posttranscriptional response might also exist, a series of reporter constructs using a CAT coding sequence attached to various portions of the TNF promoter and 3' untranslated region were devised and transfected into several cultured cell lines. All cells tested were found to be UV responsive, and in NIH 3T3 cells, induction was found to depend upon two general regions of the promoter. The more distal region encompassed nucleotides (nt) -1059 through -451 with respect to the cap site, while the more proximal region spanned nt -403 through -261. A negative element, blocking the UV response, was interposed (nt -451 through -403). As with the response to LPS, the response to UV irradiation appears to involve translational activation in macrophages. However, the UV and LPS signaling pathways have little in common with one another, as indicated by three observations. First, no difference in responsiveness was observed on comparison of TNF gene induction in macrophages derived from C3H/HeN as opposed to C3H/HeJ mice. Second, cell fusion studies showed that while the LPS signaling pathway is extinguished by fusion of RAW 264.7 cells with NIH 3T3 cells, the UV signaling pathway remained intact. Finally, induction did not depend upon the NF-kappa B binding sites that are known to be required for LPS response in macrophages, since mutation of these sites did not impair the UV response.

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Selected References

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

  1. Beutler B., Brown T. A CAT reporter construct allows ultrasensitive estimation of TNF synthesis, and suggests that the TNF gene has been silenced in non-macrophage cell lines. J Clin Invest. 1991 Apr;87(4):1336–1344. doi: 10.1172/JCI115137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Browning J. L., Ngam-ek A., Lawton P., DeMarinis J., Tizard R., Chow E. P., Hession C., O'Brine-Greco B., Foley S. F., Ware C. F. Lymphotoxin beta, a novel member of the TNF family that forms a heteromeric complex with lymphotoxin on the cell surface. Cell. 1993 Mar 26;72(6):847–856. doi: 10.1016/0092-8674(93)90574-a. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. Caput D., Beutler B., Hartog K., Thayer R., Brown-Shimer S., Cerami A. Identification of a common nucleotide sequence in the 3'-untranslated region of mRNA molecules specifying inflammatory mediators. Proc Natl Acad Sci U S A. 1986 Mar;83(6):1670–1674. doi: 10.1073/pnas.83.6.1670. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chen C., Okayama H. High-efficiency transformation of mammalian cells by plasmid DNA. Mol Cell Biol. 1987 Aug;7(8):2745–2752. doi: 10.1128/mcb.7.8.2745. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chia J. K., McManus E. J. In vitro tumor necrosis factor induction assay for analysis of febrile toxicity associated with amphotericin B preparations. Antimicrob Agents Chemother. 1990 May;34(5):906–908. doi: 10.1128/aac.34.5.906. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cseh K., Beutler B. Alternative cleavage of the cachectin/tumor necrosis factor propeptide results in a larger, inactive form of secreted protein. J Biol Chem. 1989 Sep 25;264(27):16256–16260. [PubMed] [Google Scholar]
  8. Favaloro J., Treisman R., Kamen R. Transcription maps of polyoma virus-specific RNA: analysis by two-dimensional nuclease S1 gel mapping. Methods Enzymol. 1980;65(1):718–749. doi: 10.1016/s0076-6879(80)65070-8. [DOI] [PubMed] [Google Scholar]
  9. Giroir B. P., Brown T., Beutler B. Constitutive synthesis of tumor necrosis factor in the thymus. Proc Natl Acad Sci U S A. 1992 Jun 1;89(11):4864–4868. doi: 10.1073/pnas.89.11.4864. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Giroir B. P., Peppel K., Silva M., Beutler B. The biosynthesis of tumor necrosis factor during pregnancy: studies with a CAT reporter transgene and TNF inhibitors. Eur Cytokine Netw. 1992 Nov-Dec;3(6):533–538. [PubMed] [Google Scholar]
  11. 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]
  12. Han J., Brown T., Beutler B. Endotoxin-responsive sequences control cachectin/tumor necrosis factor biosynthesis at the translational level. J Exp Med. 1990 Feb 1;171(2):465–475. doi: 10.1084/jem.171.2.465. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Han J., Huez G., Beutler B. Interactive effects of the tumor necrosis factor promoter and 3'-untranslated regions. J Immunol. 1991 Mar 15;146(6):1843–1848. [PubMed] [Google Scholar]
  14. Held W., MacDonald H. R., Weissman I. L., Hess M. W., Mueller C. Genes encoding tumor necrosis factor alpha and granzyme A are expressed during development of autoimmune diabetes. Proc Natl Acad Sci U S A. 1990 Mar;87(6):2239–2243. doi: 10.1073/pnas.87.6.2239. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Jacob C. O., Aiso S., Michie S. A., McDevitt H. O., Acha-Orbea H. Prevention of diabetes in nonobese diabetic mice by tumor necrosis factor (TNF): similarities between TNF-alpha and interleukin 1. Proc Natl Acad Sci U S A. 1990 Feb;87(3):968–972. doi: 10.1073/pnas.87.3.968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Jacob C. O., Fronek Z., Lewis G. D., Koo M., Hansen J. A., McDevitt H. O. Heritable major histocompatibility complex class II-associated differences in production of tumor necrosis factor alpha: relevance to genetic predisposition to systemic lupus erythematosus. Proc Natl Acad Sci U S A. 1990 Feb;87(3):1233–1237. doi: 10.1073/pnas.87.3.1233. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Jacob C. O., McDevitt H. O. Tumour necrosis factor-alpha in murine autoimmune 'lupus' nephritis. Nature. 1988 Jan 28;331(6154):356–358. doi: 10.1038/331356a0. [DOI] [PubMed] [Google Scholar]
  18. Kawakami M., Cerami A. Studies of endotoxin-induced decrease in lipoprotein lipase activity. J Exp Med. 1981 Sep 1;154(3):631–639. doi: 10.1084/jem.154.3.631. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Knight J. G., Adams D. D. Three genes for lupus nephritis in NZB x NZW mice. J Exp Med. 1978 Jun 1;147(6):1653–1660. doi: 10.1084/jem.147.6.1653. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kotzin B. L., Palmer E. The contribution of NZW genes to lupus-like disease in (NZB x NZW)F1 mice. J Exp Med. 1987 May 1;165(5):1237–1251. doi: 10.1084/jem.165.5.1237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kruys V., Thompson P., Beutler B. Extinction of the tumor necrosis factor locus, and of genes encoding the lipopolysaccharide signaling pathway. J Exp Med. 1993 May 1;177(5):1383–1390. doi: 10.1084/jem.177.5.1383. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Köck A., Schwarz T., Kirnbauer R., Urbanski A., Perry P., Ansel J. C., Luger T. A. Human keratinocytes are a source for tumor necrosis factor alpha: evidence for synthesis and release upon stimulation with endotoxin or ultraviolet light. J Exp Med. 1990 Dec 1;172(6):1609–1614. doi: 10.1084/jem.172.6.1609. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Morrey J. D., Bourn S. M., Bunch T. D., Jackson M. K., Sidwell R. W., Barrows L. R., Daynes R. A., Rosen C. A. In vivo activation of human immunodeficiency virus type 1 long terminal repeat by UV type A (UV-A) light plus psoralen and UV-B light in the skin of transgenic mice. J Virol. 1991 Sep;65(9):5045–5051. doi: 10.1128/jvi.65.9.5045-5051.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Piguet P. F., Grau G. E., Hauser C., Vassalli P. Tumor necrosis factor is a critical mediator in hapten induced irritant and contact hypersensitivity reactions. J Exp Med. 1991 Mar 1;173(3):673–679. doi: 10.1084/jem.173.3.673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Piguet P. F. Keratinocyte-derived tumor necrosis factor and the physiopathology of the skin. Springer Semin Immunopathol. 1992;13(3-4):345–354. doi: 10.1007/BF00200533. [DOI] [PubMed] [Google Scholar]
  26. Reinertsen J. L., Klippel J. H., Johnson A. H., Steinberg A. D., Decker J. L., Mann D. L. B-lymphocyte alloantigens associated with systemic lupus erythematosus. N Engl J Med. 1978 Sep 7;299(10):515–518. doi: 10.1056/NEJM197809072991004. [DOI] [PubMed] [Google Scholar]
  27. Satoh J., Seino H., Abo T., Tanaka S., Shintani S., Ohta S., Tamura K., Sawai T., Nobunaga T., Oteki T. Recombinant human tumor necrosis factor alpha suppresses autoimmune diabetes in nonobese diabetic mice. J Clin Invest. 1989 Oct;84(4):1345–1348. doi: 10.1172/JCI114304. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Shakhov A. N., Collart M. A., Vassalli P., Nedospasov S. A., Jongeneel C. V. Kappa B-type enhancers are involved in lipopolysaccharide-mediated transcriptional activation of the tumor necrosis factor alpha gene in primary macrophages. J Exp Med. 1990 Jan 1;171(1):35–47. doi: 10.1084/jem.171.1.35. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Sherman M. L., Datta R., Hallahan D. E., Weichselbaum R. R., Kufe D. W. Regulation of tumor necrosis factor gene expression by ionizing radiation in human myeloid leukemia cells and peripheral blood monocytes. J Clin Invest. 1991 May;87(5):1794–1797. doi: 10.1172/JCI115199. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. 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]
  31. Yoshikawa T., Streilein J. W. Genetic basis of the effects of ultraviolet light B on cutaneous immunity. Evidence that polymorphism at the Tnfa and Lps loci governs susceptibility. Immunogenetics. 1990;32(6):398–405. doi: 10.1007/BF00241633. [DOI] [PubMed] [Google Scholar]

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