Home LiteratureArticle Details
PMID: 9111316 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Expression of NFAT-family proteins in normal human T cells.

Molecular and cellular biology ·Vol. 17 ·No. 5 ·1997-05-00 ·Pages 2475-84

Lyakh L, Ghosh P, Rice NR

Abstract

NFAT proteins constitute a family of transcription factors involved in mediating signal transduction. Using a panel of specific antisera in immunoprecipitation assays, we found that NFATp (135 kDa) is constitutively expressed in normal human T cells, while synthesis of NFATc (predominant form of 86 kDa) is induced by ionomycin treatment. NFAT4/x was very weakly expressed in unstimulated cells, and its level did not increase upon treatment with activating agents. NFAT3 protein was not observed under any conditions. Higher-molecular-weight species of NFATc (of 110 and 140 kDa) were also detected. In addition, translation of NFATc mRNA apparently initiates at two different AUG codons, giving rise to proteins that differ in size by 36 amino acids. Additional size heterogeneity of both NFATc and NFATp results from phosphorylation. In contrast to ionomycin treatment, exposure of cells to phorbol myristate acetate (PMA) plus anti-CD28 did not induce NFATc, indicating that under these conditions, interleukin-2 synthesis by these cells is apparently independent of NFATc. In DNA binding assays, both PMA plus anti-CD28 and PMA plus ionomycin resulted in nuclear NFAT. Surprisingly, the PMA-ionomycin-induced synthesis of NFATc that was detected by immunoprecipitation was not mirrored in the DNA binding assays: nearly all of the activity was due to NFATp. This is the first study of expression of all family members at the protein level in normal human T cells.

MeSH Terms
CD28 Antigens/immunology DNA/metabolism DNA-Binding Proteins/metabolism Electrophoresis, Polyacrylamide Gel Humans Immune Sera Jurkat Cells Lymphocyte Activation Molecular Weight NFATC Transcription Factors Nuclear Proteins/metabolism Phosphoproteins/metabolism T-Lymphocytes/metabolism Tetradecanoylphorbol Acetate/pharmacology Transcription Factors/metabolism
Chemicals
CD28 Antigens DNA-Binding Proteins Immune Sera NFATC Transcription Factors NFATC2 protein, human NFATC3 protein, human NFATC4 protein, human Nuclear Proteins Phosphoproteins Transcription Factors DNA Tetradecanoylphorbol Acetate
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Lyakh L
Molecular Basis of Carcinogenesis Laboratory, ABL-Basic Research Program, NCI-Frederick Cancer Research and Development Center, Maryland 21702-1201, USA.
Ghosh P
Rice N R
References (45)
45 references, click to expand
  1. 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
  2. Interaction of calcineurin with a domain of the transcription factor NFAT1 that controls nuclear import.
    Proc Natl Acad Sci U S A. 1996 Aug 20;93(17):8907-12 PMID: 8799126
  3. 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
  4. 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
  5. In vivo control of NF-kappa B activation by I kappa B alpha.
    EMBO J. 1993 Dec;12(12):4685-95 PMID: 8223478
  6. Isolation of the cyclosporin-sensitive T cell transcription factor NFATp.
    Science. 1993 Oct 29;262(5134):750-4 PMID: 8235597
  7. 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
  8. 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
  9. NF-AT components define a family of transcription factors targeted in T-cell activation.
    Nature. 1994 Jun 9;369(6480):497-502 PMID: 8202141
  10. 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
  11. 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
  12. Signal transmission between the plasma membrane and nucleus of T lymphocytes.
    Annu Rev Biochem. 1994;63:1045-83 PMID: 7979236
  13. 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
  14. 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
  15. 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
  16. Cyclosporin-A sensitive induction of NF-AT in murine B cells.
    Immunity. 1994 Jun;1(3):189-96 PMID: 7889407
  17. 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
  18. 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
  19. 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
  20. 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
  21. 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
  22. 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
  23. New procedures for preparation and isolation of conjugates of proteins and a synthetic copolymer of D-amino acids and immunochemical characterization of such conjugates.
    Biochemistry. 1979 Feb 20;18(4):690-3 PMID: 84683
  24. An analysis of 5'-noncoding sequences from 699 vertebrate messenger RNAs.
    Nucleic Acids Res. 1987 Oct 26;15(20):8125-48 PMID: 3313277
  25. CD28 activation pathway regulates the production of multiple T-cell-derived lymphokines/cytokines.
    Proc Natl Acad Sci U S A. 1989 Feb;86(4):1333-7 PMID: 2465550
  26. 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. 1990 Sep 15;265(26):15788-95 PMID: 2394747
  27. The precursor of NF-kappa B p50 has I kappa B-like functions.
    Cell. 1992 Oct 16;71(2):243-53 PMID: 1423592
  28. 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
  29. 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
  30. 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
  31. The architecture of the interleukin-2 promoter: a reflection of T lymphocyte activation.
    Biochim Biophys Acta. 1995 Sep 19;1263(3):181-200 PMID: 7548205
  32. 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
  33. Transcriptional regulation of the IL-2 gene.
    Curr Opin Immunol. 1995 Jun;7(3):333-42 PMID: 7546397
  34. 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
  35. 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
  36. Hyperproliferation and dysregulation of IL-4 expression in NF-ATp-deficient mice.
    Immunity. 1996 Apr;4(4):397-405 PMID: 8612134
  37. Activation of nuclear factor of activated T cells in a cyclosporin A-resistant pathway.
    J Biol Chem. 1996 Mar 29;271(13):7700-4 PMID: 8631809
  38. Calcineurin binds the transcription factor NFAT1 and reversibly regulates its activity.
    J Biol Chem. 1996 May 3;271(18):10884-91 PMID: 8631904
  39. An enhanced immune response in mice lacking the transcription factor NFAT1.
    Science. 1996 May 10;272(5263):892-5 PMID: 8629027
  40. T-cell receptor stimulation elicits an early phase of activation and a later phase of deactivation of the transcription factor NFAT1.
    Mol Cell Biol. 1996 Jul;16(7):3945-54 PMID: 8668212
  41. Recombinant NFAT1 (NFATp) is regulated by calcineurin in T cells and mediates transcription of several cytokine genes.
    Mol Cell Biol. 1996 Jul;16(7):3955-66 PMID: 8668213
  42. Role of kinases and the phosphatase calcineurin in the nuclear shuttling of transcription factor NF-AT4.
    Nature. 1996 Jul 25;382(6589):370-3 PMID: 8684469
  43. Mechanisms of transactivation by nuclear factor of activated T cells-1.
    J Exp Med. 1996 Jul 1;184(1):141-7 PMID: 8691127
  44. Characterization of a new isoform of the NFAT (nuclear factor of activated T cells) gene family member NFATc.
    J Biol Chem. 1996 Aug 23;271(34):20914-21 PMID: 8702849
  45. 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
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.17.5.2475
Published
1997-05-00
Pages
2475-84
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC232096
Subset
IM
External Links
PubMed source DOI: 10.1128/MCB.17.5.2475 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2