The p53 tumour suppressor protein is phosphorylated at serine 389 by casein kinase II

D. W. Meek, S. Simon, U. Kikkawa, W. Eckhart

    Research output: Contribution to journalArticle

    245 Citations (Scopus)

    Abstract

    The entire coding sequence of wild-type mouse p53 was expressed in Escherichia coli under control of the PL promoter of bacteriophage lambda. The bacterial p53 protein had identical mobility to p53 from SV3T3 cells on SDS polyacrylamide gels and was recognized in bacterial lysates by three p53-specific monoclonal antibodies, including PAb246 which is specific for wild-type mouse p53. Immunoprecipitates of the bacterial p53 were phosphorylated by a highly purified preparation of rat casein kinase II; the stoichiometry of incorporation was approximately 1 mol of phosphate per mol of p53. The phosphorylated residue was identified by phosphopeptide mapping as serine 389, which is a major site of p53 phosphorylation in vivo. p53 (serine 389) kinase activity was detected on lysates of SV3T3 cells; this activity co-purified with casein kinase II on phosphocellulose and Mono Q columns and was inhibited by heparin. Immunoprecipitates of the p53-T antigen complex from SV3T3 cells also had associated serine 389 kinase activity. Phosphorylation of serine 389 by this kinase was potently inhibited by heparin and quenched by excess unlabelled GTP. The data indicate that p53 is a physiological substrate of casein kinase II, which is stimulated in response to mitogens, phosphorylates nuclear oncoproteins, and may play a role in the transduction of extracellular signals to the nucleus.
    Original languageEnglish
    Pages (from-to)3253-3260
    Number of pages8
    JournalEMBO Journal
    Volume9
    Issue number10
    Publication statusPublished - 1990

    Fingerprint

    Tumor Suppressor Protein p53
    Casein Kinase II
    Protein-Serine-Threonine Kinases
    Serine
    Phosphorylation
    Phosphotransferases
    Heparin
    Bacteriophage lambda
    Phosphopeptides
    Bacterial Proteins
    Oncogene Proteins
    Viral Tumor Antigens
    Guanosine Triphosphate
    Mitogens
    Bacteriophages
    Signal Transduction
    Phosphates
    Monoclonal Antibodies
    Stoichiometry
    Escherichia coli

    Cite this

    Meek, D. W., Simon, S., Kikkawa, U., & Eckhart, W. (1990). The p53 tumour suppressor protein is phosphorylated at serine 389 by casein kinase II. EMBO Journal, 9(10), 3253-3260.
    Meek, D. W. ; Simon, S. ; Kikkawa, U. ; Eckhart, W. / The p53 tumour suppressor protein is phosphorylated at serine 389 by casein kinase II. In: EMBO Journal. 1990 ; Vol. 9, No. 10. pp. 3253-3260.
    @article{c74c44ed52e246658852054bc33757ad,
    title = "The p53 tumour suppressor protein is phosphorylated at serine 389 by casein kinase II",
    abstract = "The entire coding sequence of wild-type mouse p53 was expressed in Escherichia coli under control of the PL promoter of bacteriophage lambda. The bacterial p53 protein had identical mobility to p53 from SV3T3 cells on SDS polyacrylamide gels and was recognized in bacterial lysates by three p53-specific monoclonal antibodies, including PAb246 which is specific for wild-type mouse p53. Immunoprecipitates of the bacterial p53 were phosphorylated by a highly purified preparation of rat casein kinase II; the stoichiometry of incorporation was approximately 1 mol of phosphate per mol of p53. The phosphorylated residue was identified by phosphopeptide mapping as serine 389, which is a major site of p53 phosphorylation in vivo. p53 (serine 389) kinase activity was detected on lysates of SV3T3 cells; this activity co-purified with casein kinase II on phosphocellulose and Mono Q columns and was inhibited by heparin. Immunoprecipitates of the p53-T antigen complex from SV3T3 cells also had associated serine 389 kinase activity. Phosphorylation of serine 389 by this kinase was potently inhibited by heparin and quenched by excess unlabelled GTP. The data indicate that p53 is a physiological substrate of casein kinase II, which is stimulated in response to mitogens, phosphorylates nuclear oncoproteins, and may play a role in the transduction of extracellular signals to the nucleus.",
    author = "Meek, {D. W.} and S. Simon and U. Kikkawa and W. Eckhart",
    year = "1990",
    language = "English",
    volume = "9",
    pages = "3253--3260",
    journal = "EMBO Journal",
    issn = "0261-4189",
    publisher = "EMBO Press",
    number = "10",

    }

    Meek, DW, Simon, S, Kikkawa, U & Eckhart, W 1990, 'The p53 tumour suppressor protein is phosphorylated at serine 389 by casein kinase II', EMBO Journal, vol. 9, no. 10, pp. 3253-3260.

    The p53 tumour suppressor protein is phosphorylated at serine 389 by casein kinase II. / Meek, D. W.; Simon, S.; Kikkawa, U.; Eckhart, W.

    In: EMBO Journal, Vol. 9, No. 10, 1990, p. 3253-3260.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - The p53 tumour suppressor protein is phosphorylated at serine 389 by casein kinase II

    AU - Meek, D. W.

    AU - Simon, S.

    AU - Kikkawa, U.

    AU - Eckhart, W.

    PY - 1990

    Y1 - 1990

    N2 - The entire coding sequence of wild-type mouse p53 was expressed in Escherichia coli under control of the PL promoter of bacteriophage lambda. The bacterial p53 protein had identical mobility to p53 from SV3T3 cells on SDS polyacrylamide gels and was recognized in bacterial lysates by three p53-specific monoclonal antibodies, including PAb246 which is specific for wild-type mouse p53. Immunoprecipitates of the bacterial p53 were phosphorylated by a highly purified preparation of rat casein kinase II; the stoichiometry of incorporation was approximately 1 mol of phosphate per mol of p53. The phosphorylated residue was identified by phosphopeptide mapping as serine 389, which is a major site of p53 phosphorylation in vivo. p53 (serine 389) kinase activity was detected on lysates of SV3T3 cells; this activity co-purified with casein kinase II on phosphocellulose and Mono Q columns and was inhibited by heparin. Immunoprecipitates of the p53-T antigen complex from SV3T3 cells also had associated serine 389 kinase activity. Phosphorylation of serine 389 by this kinase was potently inhibited by heparin and quenched by excess unlabelled GTP. The data indicate that p53 is a physiological substrate of casein kinase II, which is stimulated in response to mitogens, phosphorylates nuclear oncoproteins, and may play a role in the transduction of extracellular signals to the nucleus.

    AB - The entire coding sequence of wild-type mouse p53 was expressed in Escherichia coli under control of the PL promoter of bacteriophage lambda. The bacterial p53 protein had identical mobility to p53 from SV3T3 cells on SDS polyacrylamide gels and was recognized in bacterial lysates by three p53-specific monoclonal antibodies, including PAb246 which is specific for wild-type mouse p53. Immunoprecipitates of the bacterial p53 were phosphorylated by a highly purified preparation of rat casein kinase II; the stoichiometry of incorporation was approximately 1 mol of phosphate per mol of p53. The phosphorylated residue was identified by phosphopeptide mapping as serine 389, which is a major site of p53 phosphorylation in vivo. p53 (serine 389) kinase activity was detected on lysates of SV3T3 cells; this activity co-purified with casein kinase II on phosphocellulose and Mono Q columns and was inhibited by heparin. Immunoprecipitates of the p53-T antigen complex from SV3T3 cells also had associated serine 389 kinase activity. Phosphorylation of serine 389 by this kinase was potently inhibited by heparin and quenched by excess unlabelled GTP. The data indicate that p53 is a physiological substrate of casein kinase II, which is stimulated in response to mitogens, phosphorylates nuclear oncoproteins, and may play a role in the transduction of extracellular signals to the nucleus.

    M3 - Article

    VL - 9

    SP - 3253

    EP - 3260

    JO - EMBO Journal

    JF - EMBO Journal

    SN - 0261-4189

    IS - 10

    ER -

    Meek DW, Simon S, Kikkawa U, Eckhart W. The p53 tumour suppressor protein is phosphorylated at serine 389 by casein kinase II. EMBO Journal. 1990;9(10):3253-3260.