XCL100, an inducible nuclear MAP kinase phosphatase from Xenopus laevis: its role in MAP kinase inactivation in differentiated cells and its expression during early development

T Lewis, L A Groom, A A Sneddon, C Smythe, S M Keyse

    Research output: Contribution to journalArticle

    31 Citations (Scopus)

    Abstract

    We have cloned the Xenopus laevis homologue (XCL100) of the human CL100 (Thr/Tyr) MAP kinase phosphatase. Expression of the XCL100 mRNA and protein is inducible by serum stimulation and oxidative/heat stress in a X. laevis kidney cell line. In contrast, XCL100 is constitutively expressed in growing Xenopus oocytes. Recombinant XCL100 protein is able to dephosphorylate both tyrosine and threonine residues of activated p42 MAP kinase in vitro and both the Xenopus and human CL100 proteins were localised predominantly in the nucleus in transfected COS-1 cells. As nuclear translocation of activated MAP kinase is necessary for some of its essential functions in proliferation and cell differentiation our results indicate a role for CL100 in the regulation of these nuclear signalling events. In Xenopus kidney cells both heat shock and serum stimulation lead to transient activation of MAP kinase. However, in contrast to results previously reported from studies on mammalian fibroblasts the inactivation of MAP kinase in these epitheloid cells is rapid and is not dependent on synthesis of new protein. These results indicate that the induction of CL100 (or CL100-like enzymes) may not be required for MAP kinase inactivation in all cell types. Finally, during early embryogenesis, levels of XCL100 mRNA are greatly increased at the mid-blastula transition, suggesting that this enzyme may be involved in the regulation of MAP kinase activity during early development.
    Original languageEnglish
    Pages (from-to)2885-96
    Number of pages12
    JournalJournal of Cell Science
    Volume108 ( Pt 8)
    Publication statusPublished - 1995

    Fingerprint

    Xenopus laevis
    Phosphoric Monoester Hydrolases
    Phosphotransferases
    Xenopus
    Hot Temperature
    Blastula
    Kidney
    Messenger RNA
    COS Cells
    Mitogen-Activated Protein Kinase 1
    Enzymes
    Threonine
    Serum
    Recombinant Proteins
    Oocytes
    Embryonic Development
    Tyrosine
    Cell Differentiation
    Shock
    Proteins

    Keywords

    • Amino Acid Sequence
    • Animals
    • Base Sequence
    • Calcium-Calmodulin-Dependent Protein Kinases
    • Cell Cycle Proteins
    • Cell Differentiation
    • Cloning, Molecular
    • DNA Primers
    • Dual Specificity Phosphatase 1
    • Embryo, Nonmammalian
    • Enzyme Activation
    • Enzyme Induction
    • Female
    • Humans
    • Immediate-Early Proteins
    • Kidney
    • Kinetics
    • Mitogen-Activated Protein Kinase Phosphatases
    • Molecular Sequence Data
    • Phosphoprotein Phosphatases
    • Polymerase Chain Reaction
    • Protein Biosynthesis
    • Protein Phosphatase 1
    • Protein Tyrosine Phosphatases
    • Rabbits
    • Recombinant Proteins
    • Reticulocytes
    • Sequence Homology, Amino Acid
    • Xenopus Proteins
    • Xenopus laevis

    Cite this

    @article{cfab401c055f4b59b8dd5b18656a16b6,
    title = "XCL100, an inducible nuclear MAP kinase phosphatase from Xenopus laevis: its role in MAP kinase inactivation in differentiated cells and its expression during early development",
    abstract = "We have cloned the Xenopus laevis homologue (XCL100) of the human CL100 (Thr/Tyr) MAP kinase phosphatase. Expression of the XCL100 mRNA and protein is inducible by serum stimulation and oxidative/heat stress in a X. laevis kidney cell line. In contrast, XCL100 is constitutively expressed in growing Xenopus oocytes. Recombinant XCL100 protein is able to dephosphorylate both tyrosine and threonine residues of activated p42 MAP kinase in vitro and both the Xenopus and human CL100 proteins were localised predominantly in the nucleus in transfected COS-1 cells. As nuclear translocation of activated MAP kinase is necessary for some of its essential functions in proliferation and cell differentiation our results indicate a role for CL100 in the regulation of these nuclear signalling events. In Xenopus kidney cells both heat shock and serum stimulation lead to transient activation of MAP kinase. However, in contrast to results previously reported from studies on mammalian fibroblasts the inactivation of MAP kinase in these epitheloid cells is rapid and is not dependent on synthesis of new protein. These results indicate that the induction of CL100 (or CL100-like enzymes) may not be required for MAP kinase inactivation in all cell types. Finally, during early embryogenesis, levels of XCL100 mRNA are greatly increased at the mid-blastula transition, suggesting that this enzyme may be involved in the regulation of MAP kinase activity during early development.",
    keywords = "Amino Acid Sequence, Animals, Base Sequence, Calcium-Calmodulin-Dependent Protein Kinases, Cell Cycle Proteins, Cell Differentiation, Cloning, Molecular, DNA Primers, Dual Specificity Phosphatase 1, Embryo, Nonmammalian, Enzyme Activation, Enzyme Induction, Female, Humans, Immediate-Early Proteins, Kidney, Kinetics, Mitogen-Activated Protein Kinase Phosphatases, Molecular Sequence Data, Phosphoprotein Phosphatases, Polymerase Chain Reaction, Protein Biosynthesis, Protein Phosphatase 1, Protein Tyrosine Phosphatases, Rabbits, Recombinant Proteins, Reticulocytes, Sequence Homology, Amino Acid, Xenopus Proteins, Xenopus laevis",
    author = "T Lewis and Groom, {L A} and Sneddon, {A A} and C Smythe and Keyse, {S M}",
    year = "1995",
    language = "English",
    volume = "108 ( Pt 8)",
    pages = "2885--96",
    journal = "Journal of Cell Science",
    issn = "0021-9533",
    publisher = "Company of Biologists Ltd",

    }

    TY - JOUR

    T1 - XCL100, an inducible nuclear MAP kinase phosphatase from Xenopus laevis: its role in MAP kinase inactivation in differentiated cells and its expression during early development

    AU - Lewis, T

    AU - Groom, L A

    AU - Sneddon, A A

    AU - Smythe, C

    AU - Keyse, S M

    PY - 1995

    Y1 - 1995

    N2 - We have cloned the Xenopus laevis homologue (XCL100) of the human CL100 (Thr/Tyr) MAP kinase phosphatase. Expression of the XCL100 mRNA and protein is inducible by serum stimulation and oxidative/heat stress in a X. laevis kidney cell line. In contrast, XCL100 is constitutively expressed in growing Xenopus oocytes. Recombinant XCL100 protein is able to dephosphorylate both tyrosine and threonine residues of activated p42 MAP kinase in vitro and both the Xenopus and human CL100 proteins were localised predominantly in the nucleus in transfected COS-1 cells. As nuclear translocation of activated MAP kinase is necessary for some of its essential functions in proliferation and cell differentiation our results indicate a role for CL100 in the regulation of these nuclear signalling events. In Xenopus kidney cells both heat shock and serum stimulation lead to transient activation of MAP kinase. However, in contrast to results previously reported from studies on mammalian fibroblasts the inactivation of MAP kinase in these epitheloid cells is rapid and is not dependent on synthesis of new protein. These results indicate that the induction of CL100 (or CL100-like enzymes) may not be required for MAP kinase inactivation in all cell types. Finally, during early embryogenesis, levels of XCL100 mRNA are greatly increased at the mid-blastula transition, suggesting that this enzyme may be involved in the regulation of MAP kinase activity during early development.

    AB - We have cloned the Xenopus laevis homologue (XCL100) of the human CL100 (Thr/Tyr) MAP kinase phosphatase. Expression of the XCL100 mRNA and protein is inducible by serum stimulation and oxidative/heat stress in a X. laevis kidney cell line. In contrast, XCL100 is constitutively expressed in growing Xenopus oocytes. Recombinant XCL100 protein is able to dephosphorylate both tyrosine and threonine residues of activated p42 MAP kinase in vitro and both the Xenopus and human CL100 proteins were localised predominantly in the nucleus in transfected COS-1 cells. As nuclear translocation of activated MAP kinase is necessary for some of its essential functions in proliferation and cell differentiation our results indicate a role for CL100 in the regulation of these nuclear signalling events. In Xenopus kidney cells both heat shock and serum stimulation lead to transient activation of MAP kinase. However, in contrast to results previously reported from studies on mammalian fibroblasts the inactivation of MAP kinase in these epitheloid cells is rapid and is not dependent on synthesis of new protein. These results indicate that the induction of CL100 (or CL100-like enzymes) may not be required for MAP kinase inactivation in all cell types. Finally, during early embryogenesis, levels of XCL100 mRNA are greatly increased at the mid-blastula transition, suggesting that this enzyme may be involved in the regulation of MAP kinase activity during early development.

    KW - Amino Acid Sequence

    KW - Animals

    KW - Base Sequence

    KW - Calcium-Calmodulin-Dependent Protein Kinases

    KW - Cell Cycle Proteins

    KW - Cell Differentiation

    KW - Cloning, Molecular

    KW - DNA Primers

    KW - Dual Specificity Phosphatase 1

    KW - Embryo, Nonmammalian

    KW - Enzyme Activation

    KW - Enzyme Induction

    KW - Female

    KW - Humans

    KW - Immediate-Early Proteins

    KW - Kidney

    KW - Kinetics

    KW - Mitogen-Activated Protein Kinase Phosphatases

    KW - Molecular Sequence Data

    KW - Phosphoprotein Phosphatases

    KW - Polymerase Chain Reaction

    KW - Protein Biosynthesis

    KW - Protein Phosphatase 1

    KW - Protein Tyrosine Phosphatases

    KW - Rabbits

    KW - Recombinant Proteins

    KW - Reticulocytes

    KW - Sequence Homology, Amino Acid

    KW - Xenopus Proteins

    KW - Xenopus laevis

    M3 - Article

    VL - 108 ( Pt 8)

    SP - 2885

    EP - 2896

    JO - Journal of Cell Science

    JF - Journal of Cell Science

    SN - 0021-9533

    ER -