Cloning and Characterization of NF-ATc and NF-ATp: The Cytoplasmic Components of NF-AT

  • Steffan Ho
  • Luika Timmerman
  • Jeffrey Northrop
  • Gerald R. Crabtree

Abstract

Signalling through the T cell antigen receptor initiates a complex series of events resulting in the activation of a group of genes (early genes) that are involved in the proliferation of T cells, the development of immune function and a variety of cellular interactions that contribute to immune responses1,2. In addition, signalling through the antigen receptor is essential for several transitions in thymic development. These findings raise the question of how such diverse responses are brought about by the actions of a single receptor. Several years ago, we identified nuclear response elements for these signals in the IL-2 promoter/enhancer3–6. In the IL-2 genes, two such sites were identified that bound a protein complex that we called NF-AT after the selective expression of this complex in nuclear extracts of activated T cells5. The sequence to which this protein bound was shown to direct transcription to activated T cells in the context of a transgenic mouse7. Furthermore, transcription directed by the NF-AT binding sites required proper presentation of antigen by MHC matched cells8. These finding indicated that this protein might serve as a general terminus for signals coming from the antigen receptor and thereby aid in the elucidation of the signalling pathway carrying information from the cell membrane to the nucleus. Despite many efforts to characterize this protein, it has been elusive; indeed, it has been inferred to be a Pu protein, ets-1, elk-1, a 28 kDa protein and a 57 kDa protein. None of these proteins appear to be actually involved in the biologic activity attributed to this complex. Direct purification of the protein from bovine thymus has revealed that it is composed of two cytosolic components, NF-ATc and NF-ATp, which bear a vague similarity to a limited region of the rel/Dorsal family, but it is otherwise a pioneer protein9,10.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    G.R. Crabtree. Contingent genetic regulatory events in T lymphocyte activation. Science 243:355 (1989).PubMedCrossRefGoogle Scholar
  2. 2.
    A. Weiss. T cell antigen receptor signal transduction: a tale of tails and cytoplasmic protein-tyrosine kinases. Cell 73:209 (1993).PubMedCrossRefGoogle Scholar
  3. 3.
    U. Siebenlist, D.B. Durand, P. Bressler, N.J. Holbrook, C.A. Norris, M. Kamoun, J.A. Kant, and G.R. Crabtree. Promoter region of the IL-2 gene undergoes chromatin structure changes and confers inducibility on chloramphenicol acetyltransferase gene during activation of T cells. Mol. Cell. Biol. 6:3042 (1986).PubMedGoogle Scholar
  4. 4.
    D.B. Durand, M.R. Bush, J.G. Morgan, A. Weiss, and G.R. Crabtree. A 275 bp fragment at the 5′ end of the IL-2 gene enhances expression from a heterologous promoter in response to signals from the T cell antigen receptor. J. Exp Med. 165:395 (1987).PubMedCrossRefGoogle Scholar
  5. 5.
    J.-P. Shaw, P.J Utz, D.B. Durand, J.J. Toole, E.A. Emmel, and G.R. Crabtree. Identification of a putative regulator of early T cell activation genes. Science 241:202 (1988).PubMedCrossRefGoogle Scholar
  6. 6.
    D.B. Durand, J.-P. Shaw, M.R. Bush, R.E. Replogle, R. Belageje, and G.R. Crabtree. Characterization of antigen receptor response elements within the interleukin 2 enhancer. Mol. Cell Biol. 8:1715 (1988).PubMedGoogle Scholar
  7. 7.
    C.L. Verweij, C. Guidos, and G.R. Crabtree. Cell type specificity and activation requirements for NFAT-1 (nuclear factor of activated T-cells) transcriptional activity determined by a new method using transgenic mice to assay transcriptional activity of an individual nuclear factor. J. Biol Chem. 265:5788 (1990).Google Scholar
  8. 8.
    J. Karttunen, and N. Shastri. Measurement of ligand-induced activation in single viable T cells using the lacZ reporter gene. Proc. Natl Acad. Sci. USA 88:3972 (1991).PubMedCrossRefGoogle Scholar
  9. 9.
    J.P. Northrop, S.N. Ho, D.J. Thomas, L. Chen, G.P. Nolan, A. Admon, and G.R. Crabtree. Signaling pathways for T cell activation converge on NF-ATc, a Dorsal/Rel homologue. Nature, in press (1994).Google Scholar
  10. 10.
    P.G. McCaffrey, C. Luo, T.K. Kerppola, J. Jain, T.M. Badalian, A.M. Ho, E. Burgeon, W.S. Lane, J.N. Lambert, T. Curran, G.L. Verdine, A. Rao, and P.G. Hogan. Isolation of the cyclosporin-sensitive T cell transcription factor NFATp. Science 262:750 (1993).PubMedCrossRefGoogle Scholar
  11. 11.
    W.F. Flanagan, B. Corthesy, R.J. Bram, and G.R. Crabtree. Nuclear association of a T-cell transcription factor blocked by FK-506 and cyclosporin A. Nature 352:803 (1991).PubMedCrossRefGoogle Scholar
  12. 12.
    J.P. Northrop, K.S. Ullman, and G.R. Crabtree. Characterization of the nuclear and cytoplasmic components of the lymphoid-specific nuclear factor of activated T cells (NF-AT) complex. J. Biol Chem. 268:2917 (1993).PubMedGoogle Scholar
  13. 13.
    R.F. Doolittle. Similar amino acid sequences: chance or common ancestry. Science 214:149 (1981).PubMedCrossRefGoogle Scholar
  14. 14.
    S. Ghosh, A.M. Gifford, L.R. Riviere, P. Tempst, G.P. Nolan, and D. Baltimore. Cloning of the p50 DNA binding subunit of NF-KB: Homoloev to rel and dorsal. Cell 62:1019 (1990).PubMedCrossRefGoogle Scholar
  15. 15.
    G.P. Nolan, S. Ghosh, H.-C. Liou, P. Tempst, and D. Baltimore. DNA binding and IkB inhibition of the cloned p65 subunit of NF-κB, a rel-related polypeptide. Cell 64:961 (1991).PubMedCrossRefGoogle Scholar
  16. 16.
    G.P. Nolan, and D. Baltimore. The inhibitory ankyrin and activator Rel proteins. Current Biology, Ltd. 2:211 (1992).CrossRefGoogle Scholar
  17. 17.
    H.C. Liou, and D. Baltimore. Regulation of the NF-kappa B/rel transcription factor and I kappa B inhibitor system. Curr. Opin. Cell Biol. 5:477 (1993).PubMedCrossRefGoogle Scholar
  18. 18.
    R. Chiu, W.J. Boyle, J. Meek, T. Smeal, T. Hunter, and M. Karin, M. The c-Fos protein interacts with c-Jun/AP-1 to stimulate transcription of AP-1 responsive genes. Cell 54:541 (1988).PubMedCrossRefGoogle Scholar
  19. 19.
    T.J. Bos, D. Bohmann, H. Tsuchie, R. Tjian and P.K. Vogt, v-jun encodes a nuclear protein with enhancer binding properties of AP-1. Cell 52:705 (1988).PubMedCrossRefGoogle Scholar
  20. 20.
    S.L. Schreiber, and G.R. Crabtree. The mechanism of action of cyclosporin A and FK506. Immunology Today 13:136 (1992).PubMedCrossRefGoogle Scholar
  21. 21.
    J. Jain, P.G. McCaffrey, V.E. Valge-Archer, and A. Rao. Nuclear factor of activated T cells contains Fos and Jun. Nature 356:801 (1992).PubMedCrossRefGoogle Scholar
  22. 22.
    D. Chen, and E.V. Rothenberg. Molecular basis for developmental changes in interleukin-2 gene inducibility. Mol. Cell. Biol. 13:228 (1993).PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1994

Authors and Affiliations

  • Steffan Ho
    • 1
  • Luika Timmerman
    • 1
  • Jeffrey Northrop
    • 1
  • Gerald R. Crabtree
    • 1
    • 2
  1. 1.Department of PathologyStanford University School of MedicineStanfordUSA
  2. 2.Department of Developmental Biology and Pathology, Howard Hughes Medical Institute Rm B211Stanford University School of MedicineStanfordUSA

Personalised recommendations