Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1995 Dec 5;92(25):11623–11627. doi: 10.1073/pnas.92.25.11623

Inhibition of NF-AT-dependent transcription by NF-kappa B: implications for differential gene expression in T helper cell subsets.

V Casolaro 1, S N Georas 1, Z Song 1, I D Zubkoff 1, S A Abdulkadir 1, D Thanos 1, S J Ono 1
PMCID: PMC40454  PMID: 8524816

Abstract

Activation of individual CD4+ T cells results in differential lymphokine expression: interleukin 2 (IL-2) is preferentially produced by T helper type 1 (TH1) cells, which are involved in cell-mediated immune responses, whereas IL-4 is synthesized by TH2 cells, which are essential for humoral immunity. The Ca(2+)-dependent factor NF-ATp plays a key role in the inducible transcription of both these lymphokine genes. However, while IL2 expression requires the contribution of Ca(2+)- and protein kinase C-dependent signals, we report that activation of human IL4 transcription through the Ca(2+)-dependent pathway is diminished by protein kinase C stimulation in Jurkat T cells. This phenomenon is due to mutually exclusive binding of NF-ATp and NF-kappa B to the P sequence, an element located 69 bp upstream of the IL4 transcription initiation site. Human IL4 promoter-mediated transcription is downregulated in Jurkat cells stimulated with the NF-kappa B-activating cytokine tumor necrosis factor alpha and suppressed in RelA-overexpressing cells. In contrast, protein kinase C stimulation or RelA overexpression does not affect the activity of a human IL4 promoter containing a mouse P sequence, which is a higher-affinity site for NF-ATp and a lower-affinity site for RelA. Thus, competition between two general transcriptional activators, RelA and NF-ATp, mediates the inhibitory effect of protein kinase C stimulation on IL4 expression and may contribute to differential gene expression in TH cells.

Full text

PDF
11623

Images in this article

11624Fig. 1
on p.11624
11625Fig. 2
on p.11625
11625Fig. 3
on p.11625
11626Fig. 4
on p.11626
11626Fig. 5
on p.11626

Selected References

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

  1. Abe E., De Waal Malefyt R., Matsuda I., Arai K., Arai N. An 11-base-pair DNA sequence motif apparently unique to the human interleukin 4 gene confers responsiveness to T-cell activation signals. Proc Natl Acad Sci U S A. 1992 Apr 1;89(7):2864–2868. doi: 10.1073/pnas.89.7.2864. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Beg A. A., Baldwin A. S., Jr The I kappa B proteins: multifunctional regulators of Rel/NF-kappa B transcription factors. Genes Dev. 1993 Nov;7(11):2064–2070. doi: 10.1101/gad.7.11.2064. [DOI] [PubMed] [Google Scholar]
  3. Chuvpilo S., Schomberg C., Gerwig R., Heinfling A., Reeves R., Grummt F., Serfling E. Multiple closely-linked NFAT/octamer and HMG I(Y) binding sites are part of the interleukin-4 promoter. Nucleic Acids Res. 1993 Dec 11;21(24):5694–5704. doi: 10.1093/nar/21.24.5694. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Crabtree G. R., Clipstone N. A. Signal transmission between the plasma membrane and nucleus of T lymphocytes. Annu Rev Biochem. 1994;63:1045–1083. doi: 10.1146/annurev.bi.63.070194.005145. [DOI] [PubMed] [Google Scholar]
  5. Durand D. B., Shaw J. P., Bush M. R., Replogle R. E., Belagaje R., Crabtree G. R. Characterization of antigen receptor response elements within the interleukin-2 enhancer. Mol Cell Biol. 1988 Apr;8(4):1715–1724. doi: 10.1128/mcb.8.4.1715. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fujita T., Nolan G. P., Ghosh S., Baltimore D. Independent modes of transcriptional activation by the p50 and p65 subunits of NF-kappa B. Genes Dev. 1992 May;6(5):775–787. doi: 10.1101/gad.6.5.775. [DOI] [PubMed] [Google Scholar]
  7. Grilli M., Chiu J. J., Lenardo M. J. NF-kappa B and Rel: participants in a multiform transcriptional regulatory system. Int Rev Cytol. 1993;143:1–62. doi: 10.1016/s0074-7696(08)61873-2. [DOI] [PubMed] [Google Scholar]
  8. Hoyos B., Ballard D. W., Böhnlein E., Siekevitz M., Greene W. C. Kappa B-specific DNA binding proteins: role in the regulation of human interleukin-2 gene expression. Science. 1989 Apr 28;244(4903):457–460. doi: 10.1126/science.2497518. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Jain J., McCaffrey P. G., Miner Z., Kerppola T. K., Lambert J. N., Verdine G. L., Curran T., Rao A. The T-cell transcription factor NFATp is a substrate for calcineurin and interacts with Fos and Jun. Nature. 1993 Sep 23;365(6444):352–355. doi: 10.1038/365352a0. [DOI] [PubMed] [Google Scholar]
  11. Kang S. M., Tran A. C., Grilli M., Lenardo M. J. NF-kappa B subunit regulation in nontransformed CD4+ T lymphocytes. Science. 1992 Jun 5;256(5062):1452–1456. doi: 10.1126/science.1604322. [DOI] [PubMed] [Google Scholar]
  12. Kubo M., Kincaid R. L., Ransom J. T. Activation of the interleukin-4 gene is controlled by the unique calcineurin-dependent transcriptional factor NF(P). J Biol Chem. 1994 Jul 29;269(30):19441–19446. [PubMed] [Google Scholar]
  13. Lederer J. A., Liou J. S., Todd M. D., Glimcher L. H., Lichtman A. H. Regulation of cytokine gene expression in T helper cell subsets. J Immunol. 1994 Jan 1;152(1):77–86. [PubMed] [Google Scholar]
  14. Lenardo M. J., Fan C. M., Maniatis T., Baltimore D. The involvement of NF-kappa B in beta-interferon gene regulation reveals its role as widely inducible mediator of signal transduction. Cell. 1989 Apr 21;57(2):287–294. doi: 10.1016/0092-8674(89)90966-5. [DOI] [PubMed] [Google Scholar]
  15. Li Y. C., Ross J., Scheppler J. A., Franza B. R., Jr An in vitro transcription analysis of early responses of the human immunodeficiency virus type 1 long terminal repeat to different transcriptional activators. Mol Cell Biol. 1991 Apr;11(4):1883–1893. doi: 10.1128/mcb.11.4.1883. [DOI] [PMC free article] [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. Matsuda I., Masuda E. S., Tsuboi A., Behnam S., Arai N., Arai K. Characterization of NF(P), the nuclear factor that interacts with the regulatory P sequence (5'-CGAAAATTTCC-3') of the human interleukin-4 gene: relationship to NF-kappa B and NF-AT. Biochem Biophys Res Commun. 1994 Mar 15;199(2):439–446. doi: 10.1006/bbrc.1994.1248. [DOI] [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. Mercurio F., Karin M. Transcription factors AP-3 and AP-2 interact with the SV40 enhancer in a mutually exclusive manner. EMBO J. 1989 May;8(5):1455–1460. doi: 10.1002/j.1460-2075.1989.tb03528.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Mosmann T. R., Coffman R. L. TH1 and TH2 cells: different patterns of lymphokine secretion lead to different functional properties. Annu Rev Immunol. 1989;7:145–173. doi: 10.1146/annurev.iy.07.040189.001045. [DOI] [PubMed] [Google Scholar]
  22. Muñoz E., Zubiaga A. M., Muñoz J., Huber B. T. Regulation of IL-4 lymphokine gene expression and cellular proliferation in murine T helper type II cells. Cell Regul. 1990 Apr;1(5):425–434. doi: 10.1091/mbc.1.5.425. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Neumann M., Grieshammer T., Chuvpilo S., Kneitz B., Lohoff M., Schimpl A., Franza B. R., Jr, Serfling E. RelA/p65 is a molecular target for the immunosuppressive action of protein kinase A. EMBO J. 1995 May 1;14(9):1991–2004. doi: 10.1002/j.1460-2075.1995.tb07191.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Ono S. J., Bazil V., Sugawara M., Strominger J. L. An isotype-specific trans-acting factor is defective in a mutant B cell line that expresses HLA-DQ, but not -DR or -DP. J Exp Med. 1991 Mar 1;173(3):629–637. doi: 10.1084/jem.173.3.629. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Paul W. E., Seder R. A. Lymphocyte responses and cytokines. Cell. 1994 Jan 28;76(2):241–251. doi: 10.1016/0092-8674(94)90332-8. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. Romagnani S. Human TH1 and TH2 subsets: doubt no more. Immunol Today. 1991 Aug;12(8):256–257. doi: 10.1016/0167-5699(91)90120-I. [DOI] [PubMed] [Google Scholar]
  28. Rooney J. W., Hodge M. R., McCaffrey P. G., Rao A., Glimcher L. H. A common factor regulates both Th1- and Th2-specific cytokine gene expression. EMBO J. 1994 Feb 1;13(3):625–633. doi: 10.1002/j.1460-2075.1994.tb06300.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Rooney J. W., Hoey T., Glimcher L. H. Coordinate and cooperative roles for NF-AT and AP-1 in the regulation of the murine IL-4 gene. Immunity. 1995 May;2(5):473–483. doi: 10.1016/1074-7613(95)90028-4. [DOI] [PubMed] [Google Scholar]
  30. Sen R., Baltimore D. Inducibility of kappa immunoglobulin enhancer-binding protein Nf-kappa B by a posttranslational mechanism. Cell. 1986 Dec 26;47(6):921–928. doi: 10.1016/0092-8674(86)90807-x. [DOI] [PubMed] [Google Scholar]
  31. Szabo S. J., Gold J. S., Murphy T. L., Murphy K. M. Identification of cis-acting regulatory elements controlling interleukin-4 gene expression in T cells: roles for NF-Y and NF-ATc. Mol Cell Biol. 1993 Aug;13(8):4793–4805. doi: 10.1128/mcb.13.8.4793. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Tamura T., Yanagida T., Nariuchi H. Difference in signal transduction pathway for IL-2 and IL-4 production in T helper 1 and T helper 2 cell clones in response to anti-CD3. J Immunol. 1993 Dec 1;151(11):6051–6061. [PubMed] [Google Scholar]
  33. Tara D., Weiss D. L., Brown M. A. An activation-responsive element in the murine IL-4 gene is the site of an inducible DNA-protein interaction. J Immunol. 1993 Oct 1;151(7):3617–3626. [PubMed] [Google Scholar]
  34. Thanos D., Maniatis T. The high mobility group protein HMG I(Y) is required for NF-kappa B-dependent virus induction of the human IFN-beta gene. Cell. 1992 Nov 27;71(5):777–789. doi: 10.1016/0092-8674(92)90554-p. [DOI] [PubMed] [Google Scholar]
  35. Van der Pouw-Kraan T., Van Kooten C., Rensink I., Aarden L. Interleukin (IL)-4 production by human T cells: differential regulation of IL-4 vs. IL-2 production. Eur J Immunol. 1992 May;22(5):1237–1241. doi: 10.1002/eji.1830220519. [DOI] [PubMed] [Google Scholar]
  36. Vidard L., Colarusso L. J., Benacerraf B. Specific T-cell tolerance may reflect selective activation of lymphokine synthesis. Proc Natl Acad Sci U S A. 1995 Mar 14;92(6):2259–2262. doi: 10.1073/pnas.92.6.2259. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Ziegler-Heitbrock H. W., Wedel A., Schraut W., Ströbel M., Wendelgass P., Sternsdorf T., Bäuerle P. A., Haas J. G., Riethmüller G. Tolerance to lipopolysaccharide involves mobilization of nuclear factor kappa B with predominance of p50 homodimers. J Biol Chem. 1994 Jun 24;269(25):17001–17004. [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES