NF-ATp, a T lymphocyte DNA-binding protein that is a target for calcineurin and immunosuppressive drugs

Patricia G. McCaffrey, Brian A. Perrino, Thomas Soderling, Anjana Rao

    Research output: Contribution to journalArticle

    254 Citations (Scopus)

    Abstract

    The nuclear factor of activated T cells (NF-AT) is essential for transcription of the interleukin-2 gene upon T cell activation. Here we use a technique involving elution and renaturation of proteins from SDS-acrylamide gels to identify a DNA-binding component of NF-AT (NF-ATp) that is present in hypotonie extracts of T cells prior to activation and appears in nuclear extracts when T cells are activated. NF-ATP is present in resting T cells predominantly in a form migrating with an apparent molecular weight of 110,000-140,000, while NF-ATP from nuclear extracts of activated T cells migrates with a lower apparent molecular weight (90,000-125,000). This difference is likely to reflect dephosphorylation of NF-ATp, since treatment of NF-ATP with calf intestinal phosphatase or the calcium- and calmodulin-dependent phosphatase calcineurin in vitro results in a similar decrease in its apparent molecular weight. We show that NF-ATp is dephosphorylated in cell lysates by a calcium-dependent process that is blocked by inclusion of EGTA or a specific peptide inhibitor of calcineurin in the cell lysis buffer. Moreover, dephosphorylation of NF-ATp in cell extracts is inhibited by prior treatment of T cells with the immunosuppressive drugs cyclosporin A or FK506, which inhibit the phosphatase activity of calcineurin when complexed with their specific binding proteins, cyclophilin and FK506-binding protein. This work identifies NF-ATP as a DNA-binding phosphoprotein and a target for the drug/immunophilin/calcineurin complexes thought to mediate the inhibition of interleukin-2 gene induction by cyclosporin A and FK506.

    Original languageEnglish (US)
    Pages (from-to)3747-3752
    Number of pages6
    JournalJournal of Biological Chemistry
    Volume268
    Issue number5
    StatePublished - Feb 15 1993

    Fingerprint

    T-cells
    Calcineurin
    DNA-Binding Proteins
    Immunosuppressive Agents
    T-Lymphocytes
    Adenosine Triphosphate
    Pharmaceutical Preparations
    NFATC Transcription Factors
    Molecular Weight
    Molecular weight
    Tacrolimus
    Phosphoric Monoester Hydrolases
    Cyclosporine
    Interleukin-2
    Protein Renaturation
    Genes
    Chemical activation
    Immunophilins
    Tacrolimus Binding Proteins
    Cyclophilins

    ASJC Scopus subject areas

    • Biochemistry

    Cite this

    McCaffrey, P. G., Perrino, B. A., Soderling, T., & Rao, A. (1993). NF-ATp, a T lymphocyte DNA-binding protein that is a target for calcineurin and immunosuppressive drugs. Journal of Biological Chemistry, 268(5), 3747-3752.

    NF-ATp, a T lymphocyte DNA-binding protein that is a target for calcineurin and immunosuppressive drugs. / McCaffrey, Patricia G.; Perrino, Brian A.; Soderling, Thomas; Rao, Anjana.

    In: Journal of Biological Chemistry, Vol. 268, No. 5, 15.02.1993, p. 3747-3752.

    Research output: Contribution to journalArticle

    McCaffrey, PG, Perrino, BA, Soderling, T & Rao, A 1993, 'NF-ATp, a T lymphocyte DNA-binding protein that is a target for calcineurin and immunosuppressive drugs', Journal of Biological Chemistry, vol. 268, no. 5, pp. 3747-3752.
    McCaffrey, Patricia G. ; Perrino, Brian A. ; Soderling, Thomas ; Rao, Anjana. / NF-ATp, a T lymphocyte DNA-binding protein that is a target for calcineurin and immunosuppressive drugs. In: Journal of Biological Chemistry. 1993 ; Vol. 268, No. 5. pp. 3747-3752.
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