Skip to main content
NCBI home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1996 Oct;16(10):5232–5244. doi: 10.1128/mcb.16.10.5232

Cell-type-specific regulation of the human tumor necrosis factor alpha gene in B cells and T cells by NFATp and ATF-2/JUN.

E Y Tsai 1, J Yie 1, D Thanos 1, A E Goldfeld 1
PMCID: PMC231523  PMID: 8816436

Abstract

The human tumor necrosis factor alpha (TNF-alpha) gene is one of the earliest genes transcribed after the stimulation of a B cell through its antigen receptor or via the CD-40 pathway. In both cases, induction of TNF-alpha gene transcription can be blocked by the immunosuppressants cyclosporin A and FK506, which suggested a role for the NFAT family of proteins in the regulation of the gene in B cells. Furthermore, in T cells, two molecules of NFATp bind to the TNF-alpha promoter element kappa 3 in association with ATF-2 and Jun proteins bound to an immediately adjacent cyclic AMP response element (CRE) site. Here, using the murine B-cell lymphoma cell line A20, we show that the TNF-alpha gene is regulated in a cell-type-specific manner. In A20 B cells, the TNF-alpha gene is not regulated by NFATp bound to the kappa 3 element. Instead, ATF-2 and Jun proteins bind to the composite kappa 3/CRE site and NFATp binds to a newly identified second NFAT site centered at -76 nucleotides relative to the TNF-alpha transcription start site. This new site plays a critical role in the calcium-mediated, cyclosporin A-sensitive induction of TNF-alpha in both A20 B cells and Ar-5 cells. Consistent with these results, quantitative DNase footprinting of the TNF-alpha promoter using increasing amounts of recombinant NFATp demonstrated that the -76 site binds to NFATp with a higher affinity than the kappa 3 site. Two other previously unrecognized NFATp-binding sites in the proximal TNF-alpha promoter were also identified by this analysis. Thus, through the differential use of the same promoter element, the composite kappa 3/CRE site, the TNF-alpha gene is regulated in a cell-type-specific manner in response to the same extracellular signal.

Full Text

The Full Text of this article is available as a PDF (693.1 KB).

Selected References

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

  1. Boussiotis V. A., Nadler L. M., Strominger J. L., Goldfeld A. E. Tumor necrosis factor alpha is an autocrine growth factor for normal human B cells. Proc Natl Acad Sci U S A. 1994 Jul 19;91(15):7007–7011. doi: 10.1073/pnas.91.15.7007. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Carey M., Lin Y. S., Green M. R., Ptashne M. A mechanism for synergistic activation of a mammalian gene by GAL4 derivatives. Nature. 1990 May 24;345(6273):361–364. doi: 10.1038/345361a0. [DOI] [PubMed] [Google Scholar]
  3. Du W., Thanos D., Maniatis T. Mechanisms of transcriptional synergism between distinct virus-inducible enhancer elements. Cell. 1993 Sep 10;74(5):887–898. doi: 10.1016/0092-8674(93)90468-6. [DOI] [PubMed] [Google Scholar]
  4. Economou J. S., Rhoades K., Essner R., McBride W. H., Gasson J. C., Morton D. L. Genetic analysis of the human tumor necrosis factor alpha/cachectin promoter region in a macrophage cell line. J Exp Med. 1989 Jul 1;170(1):321–326. doi: 10.1084/jem.170.1.321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Goeddel D. V., Aggarwal B. B., Gray P. W., Leung D. W., Nedwin G. E., Palladino M. A., Patton J. S., Pennica D., Shepard H. M., Sugarman B. J. Tumor necrosis factors: gene structure and biological activities. Cold Spring Harb Symp Quant Biol. 1986;51(Pt 1):597–609. doi: 10.1101/sqb.1986.051.01.072. [DOI] [PubMed] [Google Scholar]
  6. Goldfeld A. E., Doyle C., Maniatis T. Human tumor necrosis factor alpha gene regulation by virus and lipopolysaccharide. Proc Natl Acad Sci U S A. 1990 Dec;87(24):9769–9773. doi: 10.1073/pnas.87.24.9769. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Goldfeld A. E., Flemington E. K., Boussiotis V. A., Theodos C. M., Titus R. G., Strominger J. L., Speck S. H. Transcription of the tumor necrosis factor alpha gene is rapidly induced by anti-immunoglobulin and blocked by cyclosporin A and FK506 in human B cells. Proc Natl Acad Sci U S A. 1992 Dec 15;89(24):12198–12201. doi: 10.1073/pnas.89.24.12198. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Goldfeld A. E., McCaffrey P. G., Strominger J. L., Rao A. Identification of a novel cyclosporin-sensitive element in the human tumor necrosis factor alpha gene promoter. J Exp Med. 1993 Oct 1;178(4):1365–1379. doi: 10.1084/jem.178.4.1365. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Goldfeld A. E., Strominger J. L., Doyle C. Human tumor necrosis factor alpha gene regulation in phorbol ester stimulated T and B cell lines. J Exp Med. 1991 Jul 1;174(1):73–81. doi: 10.1084/jem.174.1.73. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Goldfeld A. E., Tsai E., Kincaid R., Belshaw P. J., Schrieber S. L., Strominger J. L., Rao A. Calcineurin mediates human tumor necrosis factor alpha gene induction in stimulated T and B cells. J Exp Med. 1994 Aug 1;180(2):763–768. doi: 10.1084/jem.180.2.763. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hoey T., Sun Y. L., Williamson K., Xu X. Isolation of two new members of the NF-AT gene family and functional characterization of the NF-AT proteins. Immunity. 1995 May;2(5):461–472. doi: 10.1016/1074-7613(95)90027-6. [DOI] [PubMed] [Google Scholar]
  12. Jain J., Burgeon E., Badalian T. M., Hogan P. G., Rao A. A similar DNA-binding motif in NFAT family proteins and the Rel homology region. J Biol Chem. 1995 Feb 24;270(8):4138–4145. [PubMed] [Google Scholar]
  13. Jain J., McCaffrey P. G., Valge-Archer V. E., Rao A. Nuclear factor of activated T cells contains Fos and Jun. Nature. 1992 Apr 30;356(6372):801–804. doi: 10.1038/356801a0. [DOI] [PubMed] [Google Scholar]
  14. Leitman D. C., Ribeiro R. C., Mackow E. R., Baxter J. D., West B. L. Identification of a tumor necrosis factor-responsive element in the tumor necrosis factor alpha gene. J Biol Chem. 1991 May 25;266(15):9343–9346. [PubMed] [Google Scholar]
  15. Lin Y. S., Carey M., Ptashne M., Green M. R. How different eukaryotic transcriptional activators can cooperate promiscuously. Nature. 1990 May 24;345(6273):359–361. doi: 10.1038/345359a0. [DOI] [PubMed] [Google Scholar]
  16. Masuda E. S., Naito Y., Tokumitsu H., Campbell D., Saito F., Hannum C., Arai K., Arai N. NFATx, a novel member of the nuclear factor of activated T cells family that is expressed predominantly in the thymus. Mol Cell Biol. 1995 May;15(5):2697–2706. doi: 10.1128/mcb.15.5.2697. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. McCaffrey P. G., Goldfeld A. E., Rao A. The role of NFATp in cyclosporin A-sensitive tumor necrosis factor-alpha gene transcription. J Biol Chem. 1994 Dec 2;269(48):30445–30450. [PubMed] [Google Scholar]
  18. McCaffrey P. G., Jain J., Jamieson C., Sen R., Rao A. A T cell nuclear factor resembling NF-AT binds to an NF-kappa B site and to the conserved lymphokine promoter sequence "cytokine-1". J Biol Chem. 1992 Jan 25;267(3):1864–1871. [PubMed] [Google Scholar]
  19. McCaffrey P. G., Luo C., Kerppola T. K., Jain J., Badalian T. M., Ho A. M., Burgeon E., Lane W. S., Lambert J. N., Curran T. Isolation of the cyclosporin-sensitive T cell transcription factor NFATp. Science. 1993 Oct 29;262(5134):750–754. doi: 10.1126/science.8235597. [DOI] [PubMed] [Google Scholar]
  20. Miner J. N., Yamamoto K. R. The basic region of AP-1 specifies glucocorticoid receptor activity at a composite response element. Genes Dev. 1992 Dec;6(12B):2491–2501. doi: 10.1101/gad.6.12b.2491. [DOI] [PubMed] [Google Scholar]
  21. Nolan G. P. NF-AT-AP-1 and Rel-bZIP: hybrid vigor and binding under the influence. Cell. 1994 Jun 17;77(6):795–798. doi: 10.1016/0092-8674(94)90126-0. [DOI] [PubMed] [Google Scholar]
  22. Northrop J. P., Ho S. N., Chen L., Thomas D. J., Timmerman L. A., Nolan G. P., Admon A., Crabtree G. R. NF-AT components define a family of transcription factors targeted in T-cell activation. Nature. 1994 Jun 9;369(6480):497–502. doi: 10.1038/369497a0. [DOI] [PubMed] [Google Scholar]
  23. Rao A. NF-ATp: a transcription factor required for the co-ordinate induction of several cytokine genes. Immunol Today. 1994 Jun;15(6):274–281. doi: 10.1016/0167-5699(94)90007-8. [DOI] [PubMed] [Google Scholar]
  24. Rooney J. W., Sun Y. L., Glimcher L. H., Hoey T. Novel NFAT sites that mediate activation of the interleukin-2 promoter in response to T-cell receptor stimulation. Mol Cell Biol. 1995 Nov;15(11):6299–6310. doi: 10.1128/mcb.15.11.6299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Takashiba S., Van Dyke T. E., Shapira L., Amar S. Lipopolysaccharide-inducible and salicylate-sensitive nuclear factor(s) on human tumor necrosis factor alpha promoter. Infect Immun. 1995 Apr;63(4):1529–1534. doi: 10.1128/iai.63.4.1529-1534.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Thanos D., Maniatis T. Virus induction of human IFN beta gene expression requires the assembly of an enhanceosome. Cell. 1995 Dec 29;83(7):1091–1100. doi: 10.1016/0092-8674(95)90136-1. [DOI] [PubMed] [Google Scholar]
  27. Tsai E. Y., Jain J., Pesavento P. A., Rao A., Goldfeld A. E. Tumor necrosis factor alpha gene regulation in activated T cells involves ATF-2/Jun and NFATp. Mol Cell Biol. 1996 Feb;16(2):459–467. doi: 10.1128/mcb.16.2.459. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Venkataraman L., Francis D. A., Wang Z., Liu J., Rothstein T. L., Sen R. Cyclosporin-A sensitive induction of NF-AT in murine B cells. Immunity. 1994 Jun;1(3):189–196. doi: 10.1016/1074-7613(94)90097-3. [DOI] [PubMed] [Google Scholar]
  29. Xie H., Chiles T. C., Rothstein T. L. Induction of CREB activity via the surface Ig receptor of B cells. J Immunol. 1993 Jul 15;151(2):880–889. [PubMed] [Google Scholar]
  30. Yaseen N. R., Maizel A. L., Wang F., Sharma S. Comparative analysis of NFAT (nuclear factor of activated T cells) complex in human T and B lymphocytes. J Biol Chem. 1993 Jul 5;268(19):14285–14293. [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES