Skip to main content

Some NLM-NCBI services and products are experiencing heavy traffic, which may affect performance and availability. We apologize for the inconvenience and appreciate your patience. For assistance, please contact our Help Desk at info@ncbi.nlm.nih.gov.

The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1994 Mar 1;179(3):931–942. doi: 10.1084/jem.179.3.931

Interleukin 2 transcription factors as molecular targets of cAMP inhibition: delayed inhibition kinetics and combinatorial transcription roles

PMCID: PMC2191402  PMID: 8113685

Abstract

Elevation of cAMP can cause gene-specific inhibition of interleukin 2 (IL-2) expression. To investigate the mechanism of this effect, we have combined electrophoretic mobility shift assays and in vivo genomic footprinting to assess both the availability of putative IL-2 transcription factors in forskolin-treated cells and the functional capacity of these factors to engage their sites in vivo. All observed effects of forskolin depended upon protein kinase A, for they were blocked by introduction of a dominant negative mutant subunit of protein kinase A. In the EL4.E1 cell line, we report specific inhibitory effects of cAMP elevation both on NF-kappa B/Rel family factors binding at -200 bp, and on a novel, biochemically distinct "TGGGC" factor binding at -225 bp with respect to the IL-2 transcriptional start site. Neither NF-AT nor AP-1 binding activities are detectably inhibited in gel mobility shift assays. Elevation of cAMP inhibits NF-kappa B activity with delayed kinetics in association with a delayed inhibition of IL-2 RNA accumulation. Activation of cells in the presence of forskolin prevents the maintenance of stable protein- DNA interactions in vivo, not only at the NF-kappa B and TGGGC sites of the IL-2 enhancer, but also at the NF-AT, AP-1, and other sites. This result, and similar results in cyclosporin A-treated cells, imply that individual IL-2 transcription factors cannot stably bind their target sequences in vivo without coengagement of all other distinct factors at neighboring sites. It is proposed that nonhierarchical, cooperative enhancement of binding is a structural basis of combinatorial transcription factor action at the IL-2 locus.

Full Text

The Full Text of this article is available as a PDF (1.9 MB).

Selected References

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

  1. Averill L. E., Stein R. L., Kammer G. M. Control of human T-lymphocyte interleukin-2 production by a cAMP-dependent pathway. Cell Immunol. 1988 Aug;115(1):88–99. doi: 10.1016/0008-8749(88)90164-5. [DOI] [PubMed] [Google Scholar]
  2. Baeuerle P. A., Baltimore D. Activation of DNA-binding activity in an apparently cytoplasmic precursor of the NF-kappa B transcription factor. Cell. 1988 Apr 22;53(2):211–217. doi: 10.1016/0092-8674(88)90382-0. [DOI] [PubMed] [Google Scholar]
  3. Betz M., Fox B. S. Prostaglandin E2 inhibits production of Th1 lymphokines but not of Th2 lymphokines. J Immunol. 1991 Jan 1;146(1):108–113. [PubMed] [Google Scholar]
  4. Chedid M., Mizel S. B. Involvement of cyclic AMP-dependent protein kinases in the signal transduction pathway for interleukin-1. Mol Cell Biol. 1990 Jul;10(7):3824–3827. doi: 10.1128/mcb.10.7.3824. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chen D., Rothenberg E. V. Molecular basis for developmental changes in interleukin-2 gene inducibility. Mol Cell Biol. 1993 Jan;13(1):228–237. doi: 10.1128/mcb.13.1.228. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Daynes R. A., Araneo B. A. Contrasting effects of glucocorticoids on the capacity of T cells to produce the growth factors interleukin 2 and interleukin 4. Eur J Immunol. 1989 Dec;19(12):2319–2325. doi: 10.1002/eji.1830191221. [DOI] [PubMed] [Google Scholar]
  7. Flanagan W. M., Corthésy B., Bram R. J., Crabtree G. R. Nuclear association of a T-cell transcription factor blocked by FK-506 and cyclosporin A. Nature. 1991 Aug 29;352(6338):803–807. doi: 10.1038/352803a0. [DOI] [PubMed] [Google Scholar]
  8. Gajewski T. F., Schell S. R., Fitch F. W. Evidence implicating utilization of different T cell receptor-associated signaling pathways by TH1 and TH2 clones. J Immunol. 1990 Jun 1;144(11):4110–4120. [PubMed] [Google Scholar]
  9. Garrity P. A., Wold B. J. Effects of different DNA polymerases in ligation-mediated PCR: enhanced genomic sequencing and in vivo footprinting. Proc Natl Acad Sci U S A. 1992 Feb 1;89(3):1021–1025. doi: 10.1073/pnas.89.3.1021. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Go C., Miller J. Differential induction of transcription factors that regulate the interleukin 2 gene during anergy induction and restimulation. J Exp Med. 1992 May 1;175(5):1327–1336. doi: 10.1084/jem.175.5.1327. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. Hagiwara M., Brindle P., Harootunian A., Armstrong R., Rivier J., Vale W., Tsien R., Montminy M. R. Coupling of hormonal stimulation and transcription via the cyclic AMP-responsive factor CREB is rate limited by nuclear entry of protein kinase A. Mol Cell Biol. 1993 Aug;13(8):4852–4859. doi: 10.1128/mcb.13.8.4852. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hohmann H. P., Remy R., Scheidereit C., van Loon A. P. Maintenance of NF-kappa B activity is dependent on protein synthesis and the continuous presence of external stimuli. Mol Cell Biol. 1991 Jan;11(1):259–266. doi: 10.1128/mcb.11.1.259. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kang S. M., Beverly B., Tran A. C., Brorson K., Schwartz R. H., Lenardo M. J. Transactivation by AP-1 is a molecular target of T cell clonal anergy. Science. 1992 Aug 21;257(5073):1134–1138. doi: 10.1126/science.257.5073.1134. [DOI] [PubMed] [Google Scholar]
  15. Lerner A., Jacobson B., Miller R. A. Cyclic AMP concentrations modulate both calcium flux and hydrolysis of phosphatidylinositol phosphates in mouse T lymphocytes. J Immunol. 1988 Feb 1;140(3):936–940. [PubMed] [Google Scholar]
  16. Liu Y., Janeway C. A., Jr Interferon gamma plays a critical role in induced cell death of effector T cell: a possible third mechanism of self-tolerance. J Exp Med. 1990 Dec 1;172(6):1735–1739. doi: 10.1084/jem.172.6.1735. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Mary D., Aussel C., Ferrua B., Fehlmann M. Regulation of interleukin 2 synthesis by cAMP in human T cells. J Immunol. 1987 Aug 15;139(4):1179–1184. [PubMed] [Google Scholar]
  18. McGuire K. L., Yang J. A., Rothenberg E. V. Influence of activating stimulus on functional phenotype: interleukin 2 mRNA accumulation differentially induced by ionophore and receptor ligands in subsets of murine T cells. Proc Natl Acad Sci U S A. 1988 Sep;85(17):6503–6507. doi: 10.1073/pnas.85.17.6503. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. McKnight G. S., Cadd G. G., Clegg C. H., Otten A. D., Correll L. A. Expression of wild-type and mutant subunits of the cAMP-dependent protein kinase. Cold Spring Harb Symp Quant Biol. 1988;53(Pt 1):111–119. doi: 10.1101/sqb.1988.053.01.017. [DOI] [PubMed] [Google Scholar]
  20. Mueller D. L., Jenkins M. K., Schwartz R. H. Clonal expansion versus functional clonal inactivation: a costimulatory signalling pathway determines the outcome of T cell antigen receptor occupancy. Annu Rev Immunol. 1989;7:445–480. doi: 10.1146/annurev.iy.07.040189.002305. [DOI] [PubMed] [Google Scholar]
  21. Muñoz E., Zubiaga A. M., Merrow M., Sauter N. P., Huber B. T. Cholera toxin discriminates between T helper 1 and 2 cells in T cell receptor-mediated activation: role of cAMP in T cell proliferation. J Exp Med. 1990 Jul 1;172(1):95–103. doi: 10.1084/jem.172.1.95. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Northrop J. P., Crabtree G. R., Mattila P. S. Negative regulation of interleukin 2 transcription by the glucocorticoid receptor. J Exp Med. 1992 May 1;175(5):1235–1245. doi: 10.1084/jem.175.5.1235. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Novak T. J., Chen D., Rothenberg E. V. Interleukin-1 synergy with phosphoinositide pathway agonists for induction of interleukin-2 gene expression: molecular basis of costimulation. Mol Cell Biol. 1990 Dec;10(12):6325–6334. doi: 10.1128/mcb.10.12.6325. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Novak T. J., Rothenberg E. V. Differential transient and long-term expression of DNA sequences introduced into T-lymphocyte lines. DNA. 1986 Dec;5(6):439–451. doi: 10.1089/dna.1.1986.5.439. [DOI] [PubMed] [Google Scholar]
  25. Novak T. J., Rothenberg E. V. cAMP inhibits induction of interleukin 2 but not of interleukin 4 in T cells. Proc Natl Acad Sci U S A. 1990 Dec;87(23):9353–9357. doi: 10.1073/pnas.87.23.9353. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. O'Shea J. J., Urdahl K. B., Luong H. T., Chused T. M., Samelson L. E., Klausner R. D. Aluminum fluoride induces phosphatidylinositol turnover, elevation of cytoplasmic free calcium, and phosphorylation of the T cell antigen receptor in murine T cells. J Immunol. 1987 Nov 15;139(10):3463–3469. [PubMed] [Google Scholar]
  27. Riegel J. S., Corthesy B., Flanagan W. M., Crabtree G. R. Regulation of the interleukin-2 gene. Chem Immunol. 1992;51:266–298. [PubMed] [Google Scholar]
  28. Shirakawa F., Chedid M., Suttles J., Pollok B. A., Mizel S. B. Interleukin 1 and cyclic AMP induce kappa immunoglobulin light-chain expression via activation of an NF-kappa B-like DNA-binding protein. Mol Cell Biol. 1989 Mar;9(3):959–964. doi: 10.1128/mcb.9.3.959. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Shirakawa F., Mizel S. B. In vitro activation and nuclear translocation of NF-kappa B catalyzed by cyclic AMP-dependent protein kinase and protein kinase C. Mol Cell Biol. 1989 Jun;9(6):2424–2430. doi: 10.1128/mcb.9.6.2424. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Vacca A., Felli M. P., Farina A. R., Martinotti S., Maroder M., Screpanti I., Meco D., Petrangeli E., Frati L., Gulino A. Glucocorticoid receptor-mediated suppression of the interleukin 2 gene expression through impairment of the cooperativity between nuclear factor of activated T cells and AP-1 enhancer elements. J Exp Med. 1992 Mar 1;175(3):637–646. doi: 10.1084/jem.175.3.637. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press

RESOURCES