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PMID: 8668213 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Recombinant NFAT1 (NFATp) is regulated by calcineurin in T cells and mediates transcription of several cytokine genes.

Molecular and cellular biology ·Vol. 16 ·No. 7 ·1996-07-00 ·Pages 3955-66

Luo C, Burgeon E, Carew JA, McCaffrey PG, Badalian TM, Lane WS, Hogan PG, Rao A

Abstract

Transcription factors of the NFAT family play a key role in the transcription of cytokine genes and other genes during the immune response. We have identified two new isoforms of the transcription factor NFAT1 (previously termed NFATp) that are the predominant isoforms expressed in murine and human T cells. When expressed in Jurkat T cells, recombinant NFAT1 is regulated, as expected, by the calmodulin-dependent phosphatase calcineurin, and its function is inhibited by the immunosuppressive agent cyclosporin A (CsA). Transactivation by recombinant NFAT1 in Jurkat T cells requires dual stimulation with ionomycin and phorbol 12-myristate 13-acetate; this activity is potentiated by coexpression of constitutively active calcineurin and is inhibited by CsA. Immunocytochemical analysis indicates that recombinant NFAT1 localizes in the cytoplasm of transiently transfected T cells and translocates into the nucleus in a CsA-sensitive manner following ionomycin stimulation. When expressed in COS cells, however, NFAT1 is capable of transactivation, but it is not regulated correctly: its subcellular localization and transcriptional function are not affected by stimulation of the COS cells with ionomycin and phorbol 12-myristate 13-acetate. Recombinant NFAT1 can mediate transcription of the interleukin-2, interleukin-4, tumor necrosis factor alpha, and granulocyte-macrophage colony-stimulating factor promoters in T cells, suggesting that NFAT1 contributes to the CsA-sensitive transcription of these genes during the immune response.

MeSH Terms
Amino Acid Sequence Animals Base Sequence Cell Line Chloramphenicol O-Acetyltransferase/metabolism Chlorocebus aethiops Cloning, Molecular Cytokines/biosynthesis DNA Primers DNA-Binding Proteins/biosynthesis,chemistry,metabolism Gene Expression Regulation Granulocyte-Macrophage Colony-Stimulating Factor/biosynthesis Humans Interleukin-2/biosynthesis Interleukin-4/biosynthesis Mice Molecular Sequence Data NFATC Transcription Factors Nuclear Proteins/metabolism Polymerase Chain Reaction Promoter Regions, Genetic Recombinant Proteins/biosynthesis,chemistry,metabolism Sequence Homology, Amino Acid T-Lymphocytes/physiology Transcription Factors/biosynthesis,chemistry,metabolism Transcription, Genetic Transcriptional Activation Transfection Tumor Cells, Cultured Tumor Necrosis Factor-alpha/biosynthesis
Chemicals
Cytokines DNA Primers DNA-Binding Proteins Interleukin-2 NFATC Transcription Factors NFATC2 protein, human Nfatc2 protein, mouse Nuclear Proteins Recombinant Proteins Transcription Factors Tumor Necrosis Factor-alpha Interleukin-4 Granulocyte-Macrophage Colony-Stimulating Factor Chloramphenicol O-Acetyltransferase
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Luo C
Division of Cellular and Molecular Biology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts 02115, USA.
Burgeon E
Carew J A
McCaffrey P G
Badalian T M
Lane W S
Hogan P G
Rao A
References (69)
69 references, click to expand
  1. Phosphorylation of the transcription factor NFATp inhibits its DNA binding activity in cyclosporin A-treated human B and T cells.
    J Biol Chem. 1995 Sep 1;270(35):20653-9 PMID: 7657645
  2. An ion pair in class II major histocompatibility complex heterodimers critical for surface expression and peptide presentation.
    J Biol Chem. 1995 Sep 22;270(38):22351-60 PMID: 7673219
  3. Direct demonstration of NFATp dephosphorylation and nuclear localization in activated HT-2 cells using a specific NFATp polyclonal antibody.
    J Biol Chem. 1995 Sep 22;270(38):22602-7 PMID: 7545680
  4. Only one of the two DNA-bound orientations of AP-1 found in solution cooperates with NFATp.
    Curr Biol. 1995 Aug 1;5(8):882-9 PMID: 7583146
  5. The cyclosporin-sensitive transcription factor NFATp is expressed in several classes of cells in the immune system.
    Ann N Y Acad Sci. 1995 Sep 7;766:182-94 PMID: 7486655
  6. The actions of cyclosporin A and FK506 suggest a novel step in the activation of T lymphocytes.
    EMBO J. 1990 Dec;9(13):4425-33 PMID: 1702384
  7. Critical structural elements of the VP16 transcriptional activation domain.
    Science. 1991 Jan 4;251(4989):87-90 PMID: 1846049
  8. The immunosuppressives FK 506 and cyclosporin A inhibit the generation of protein factors binding to the two purine boxes of the interleukin 2 enhancer.
    Nucleic Acids Res. 1991 Jan 11;19(1):61-7 PMID: 1707162
  9. Nuclear association of a T-cell transcription factor blocked by FK-506 and cyclosporin A.
    Nature. 1991 Aug 29;352(6338):803-7 PMID: 1715516
  10. Functional characterization of the NF-kappa B p65 transcriptional activator and an alternatively spliced derivative.
    Mol Cell Biol. 1992 Feb;12(2):444-54 PMID: 1732726
  11. Nuclear factor of activated T cells contains Fos and Jun.
    Nature. 1992 Apr 30;356(6372):801-4 PMID: 1533441
  12. FK-506- and CsA-sensitive activation of the interleukin-2 promoter by calcineurin.
    Nature. 1992 Jun 25;357(6380):692-4 PMID: 1377361
  13. p21ras and calcineurin synergize to regulate the nuclear factor of activated T cells.
    J Exp Med. 1993 Nov 1;178(5):1517-22 PMID: 8228805
  14. Isolation of the cyclosporin-sensitive T cell transcription factor NFATp.
    Science. 1993 Oct 29;262(5134):750-4 PMID: 8235597
  15. The granulocyte-macrophage colony-stimulating factor promoter cis-acting element CLE0 mediates induction signals in T cells and is recognized by factors related to AP1 and NFAT.
    Mol Cell Biol. 1993 Dec;13(12):7399-407 PMID: 8246960
  16. 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):5694-704 PMID: 8284217
  17. A common factor regulates both Th1- and Th2-specific cytokine gene expression.
    EMBO J. 1994 Feb 1;13(3):625-33 PMID: 8313907
  18. The Ca2+/calmodulin-activated, phosphoprotein phosphatase calcineurin is sufficient for positive transcriptional regulation of the mouse IL-4 gene.
    Int Immunol. 1994 Feb;6(2):179-88 PMID: 8155595
  19. Nuclear localization and cell cycle regulation of a murine protein tyrosine phosphatase.
    Mol Cell Biol. 1994 May;14(5):3030-40 PMID: 8164659
  20. Delineation of a region in the B2 bradykinin receptor that is essential for high-affinity agonist binding.
    Proc Natl Acad Sci U S A. 1994 May 10;91(10):4417-21 PMID: 8183923
  21. Calcineurin potentiates activation of the granulocyte-macrophage colony-stimulating factor gene in T cells: involvement of the conserved lymphokine element 0.
    Mol Biol Cell. 1994 Jan;5(1):119-28 PMID: 8186461
  22. NF-AT components define a family of transcription factors targeted in T-cell activation.
    Nature. 1994 Jun 9;369(6480):497-502 PMID: 8202141
  23. NF-AT-AP-1 and Rel-bZIP: hybrid vigor and binding under the influence.
    Cell. 1994 Jun 17;77(6):795-8 PMID: 8004669
  24. Calcineurin mediates human tumor necrosis factor alpha gene induction in stimulated T and B cells.
    J Exp Med. 1994 Aug 1;180(2):763-8 PMID: 8046352
  25. NF-ATp: a transcription factor required for the co-ordinate induction of several cytokine genes.
    Immunol Today. 1994 Jun;15(6):274-81 PMID: 8068174
  26. Structural and functional analysis of the NF-kappa B p65 C terminus. An acidic and modular transactivation domain with the potential to adopt an alpha-helical conformation.
    J Biol Chem. 1994 Oct 14;269(41):25613-20 PMID: 7929265
  27. The latch region of calcineurin B is involved in both immunosuppressant-immunophilin complex docking and phosphatase activation.
    Cell. 1994 Nov 4;79(3):437-47 PMID: 7525078
  28. Expression of the transcription factor NFATp in a neuronal cell line and in the murine nervous system.
    J Biol Chem. 1994 Nov 11;269(45):28181-6 PMID: 7961754
  29. The role of NFATp in cyclosporin A-sensitive tumor necrosis factor-alpha gene transcription.
    J Biol Chem. 1994 Dec 2;269(48):30445-50 PMID: 7982959
  30. Calcium regulation of calcineurin phosphatase activity by its B subunit and calmodulin. Role of the autoinhibitory domain.
    J Biol Chem. 1995 Jan 6;270(1):340-6 PMID: 7814394
  31. Structure of NF-kappa B p50 homodimer bound to a kappa B site.
    Nature. 1995 Jan 26;373(6512):303-10 PMID: 7530332
  32. Structure of the NF-kappa B p50 homodimer bound to DNA.
    Nature. 1995 Jan 26;373(6512):311-7 PMID: 7830764
  33. A similar DNA-binding motif in NFAT family proteins and the Rel homology region.
    J Biol Chem. 1995 Feb 24;270(8):4138-45 PMID: 7876165
  34. Induction of NF-AT in normal B lymphocytes by anti-immunoglobulin or CD40 ligand in conjunction with IL-4.
    Immunity. 1994 Jun;1(3):179-87 PMID: 7889406
  35. Cyclosporin-A sensitive induction of NF-AT in murine B cells.
    Immunity. 1994 Jun;1(3):189-96 PMID: 7889407
  36. Human granulocyte-macrophage colony-stimulating factor enhancer function is associated with cooperative interactions between AP-1 and NFATp/c.
    Mol Cell Biol. 1995 Apr;15(4):2071-9 PMID: 7891702
  37. 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):2697-706 PMID: 7739550
  38. Isolation of two new members of the NF-AT gene family and functional characterization of the NF-AT proteins.
    Immunity. 1995 May;2(5):461-72 PMID: 7749981
  39. Coordinate and cooperative roles for NF-AT and AP-1 in the regulation of the murine IL-4 gene.
    Immunity. 1995 May;2(5):473-83 PMID: 7749982
  40. A nuclear factor of activated T cell-like transcription factor in mast cells is involved in IL-5 gene regulation after IgE plus antigen stimulation.
    J Immunol. 1995 Jun 1;154(11):6112-9 PMID: 7751652
  41. Characterization of a mutant calcineurin A alpha gene expressed by EL4 lymphoma cells.
    Mol Cell Biol. 1995 Jul;15(7):3857-63 PMID: 7791792
  42. Fc epsilon RI-mediated induction of nuclear factor of activated T-cells.
    J Biol Chem. 1995 Jul 7;270(27):16333-8 PMID: 7608202
  43. Multiple signals are required for function of the human granulocyte-macrophage colony-stimulating factor gene promoter in T cells.
    J Immunol. 1995 Aug 1;155(3):1240-51 PMID: 7636192
  44. NFATc3, a lymphoid-specific NFATc family member that is calcium-regulated and exhibits distinct DNA binding specificity.
    J Biol Chem. 1995 Aug 25;270(34):19898-907 PMID: 7650004
  45. Immunosuppressive drugs prevent a rapid dephosphorylation of transcription factor NFAT1 in stimulated immune cells.
    Proc Natl Acad Sci U S A. 1995 Nov 21;92(24):11205-9 PMID: 7479966
  46. Identification of a physical interaction between calcineurin and nuclear factor of activated T cells (NFATp).
    J Biol Chem. 1996 Jan 19;271(3):1274-7 PMID: 8576111
  47. Hyperproliferation and dysregulation of IL-4 expression in NF-ATp-deficient mice.
    Immunity. 1996 Apr;4(4):397-405 PMID: 8612134
  48. Calcineurin binds the transcription factor NFAT1 and reversibly regulates its activity.
    J Biol Chem. 1996 May 3;271(18):10884-91 PMID: 8631904
  49. An enhanced immune response in mice lacking the transcription factor NFAT1.
    Science. 1996 May 10;272(5263):892-5 PMID: 8629027
  50. Identification of calcineurin as a key signalling enzyme in T-lymphocyte activation.
    Nature. 1992 Jun 25;357(6380):695-7 PMID: 1377362
  51. Characterization of the nuclear and cytoplasmic components of the lymphoid-specific nuclear factor of activated T cells (NF-AT) complex.
    J Biol Chem. 1993 Feb 5;268(4):2917-23 PMID: 8428966
  52. NF-ATp, a T lymphocyte DNA-binding protein that is a target for calcineurin and immunosuppressive drugs.
    J Biol Chem. 1993 Feb 15;268(5):3747-52 PMID: 7679116
  53. Both N- and C-terminal domains of RelB are required for full transactivation: role of the N-terminal leucine zipper-like motif.
    Mol Cell Biol. 1993 Mar;13(3):1572-82 PMID: 8441398
  54. The NFAT-1 DNA binding complex in activated T cells contains Fra-1 and JunB.
    Mol Cell Biol. 1993 Mar;13(3):1911-9 PMID: 8441422
  55. Transcription of the interleukin 4 gene is regulated by multiple promoter elements.
    J Exp Med. 1993 Jun 1;177(6):1663-74 PMID: 8496684
  56. 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):14285-93 PMID: 8314792
  57. Analysis of the preexisting and nuclear forms of nuclear factor of activated T cells.
    J Immunol. 1993 Jul 15;151(2):837-48 PMID: 8335913
  58. 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):4793-805 PMID: 8336717
  59. Identification of a novel cyclosporin-sensitive element in the human tumor necrosis factor alpha gene promoter.
    J Exp Med. 1993 Oct 1;178(4):1365-79 PMID: 8376940
  60. The T-cell transcription factor NFATp is a substrate for calcineurin and interacts with Fos and Jun.
    Nature. 1993 Sep 23;365(6444):352-5 PMID: 8397339
  61. FK506 and cyclosporin, molecular probes for studying intracellular signal transduction.
    Immunol Today. 1993 Jun;14(6):290-5 PMID: 7691065
  62. Promoter region of interleukin-2 gene undergoes chromatin structure changes and confers inducibility on chloramphenicol acetyltransferase gene during activation of T cells.
    Mol Cell Biol. 1986 Sep;6(9):3042-9 PMID: 3491296
  63. Identification of a putative regulator of early T cell activation genes.
    Science. 1988 Jul 8;241(4862):202-5 PMID: 3260404
  64. B-cell-specific enhancer activity of conserved upstream elements of the class II major histocompatibility complex DQB gene.
    Proc Natl Acad Sci U S A. 1988 Sep;85(18):6909-13 PMID: 3137578
  65. The immunosuppressant FK506 selectively inhibits expression of early T cell activation genes.
    J Immunol. 1989 Jul 15;143(2):718-26 PMID: 2472451
  66. Transcriptional regulation in mammalian cells by sequence-specific DNA binding proteins.
    Science. 1989 Jul 28;245(4916):371-8 PMID: 2667136
  67. 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):5169-81 PMID: 2557547
  68. pEF-BOS, a powerful mammalian expression vector.
    Nucleic Acids Res. 1990 Sep 11;18(17):5322 PMID: 1698283
  69. Activation and expression of the nuclear factors of activated T cells, NFATp and NFATc, in human natural killer cells: regulation upon CD16 ligand binding.
    J Exp Med. 1995 Sep 1;182(3):801-10 PMID: 7650486
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.16.7.3955
Published
1996-07-00
Pages
3955-66
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC231392
Subset
IM
Grants
NCI NIH HHS · CA42471 · United States
NIGMS NIH HHS · GM46227 · United States
Databases
GENBANK
U02079, U36575, U36576, U43341, U43342
External Links
PubMed source DOI: 10.1128/MCB.16.7.3955 PMC Full Text View journal details
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