Isolation of the human genes encoding the pyst1 and Pyst2 phosphatases: characterisation of Pyst2 as a cytosolic dual-specificity MAP kinase phosphatase and its catalytic activation by both MAP and SAP kinases

S Dowd, A A Sneddon, S M Keyse

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    103 Citations (Scopus)

    Abstract

    We have isolated the human genes encoding the Pyst1 (MKP-3) and Pyst2 (MKP-X) MAP kinase phosphatases. Both genes consist of three exons interrupted by two introns and lack an intron which is conserved in all the other members of this gene family characterised to date. This reinforces the conclusion that Pyst1 and Pyst2 are members of a distinct and structurally homologous subfamily of dual-specificity (Thr/Tyr) MAP kinase phosphatases. We find that Pyst2 mRNA is constitutively expressed in a wide variety of human cell lines including those derived from ovarian, bladder and breast cancers. While there is no evidence for inducible expression of Pyst2 mRNA in human skin fibroblasts in response to cellular stress, Pyst2 mRNA levels are moderately increased in response to serum stimulation. Pyst2 protein is predominantly cytosolic when expressed in COS-1 cells. In common with Pyst1, Pyst2 shows substrate selectivity for the classical p42 (ERK2) isoform of MAP kinase both in vitro and in vivo, displaying much reduced activity towards stress activated MAP kinase isoforms such as JNK-1 and p38/RK. Pyst2 binds p42 MAP kinase in vivo and both MAP kinase binding and substrate selectivity correlate with the ability of different recombinant MAP and SAP kinases to cause catalytic activation of the Pyst2 phosphatase in vitro.
    Original languageEnglish
    Pages (from-to)3389-99
    Number of pages11
    JournalJournal of Cell Science
    Volume111 ( Pt 22)
    Publication statusPublished - 1998

    Fingerprint

    Phosphoric Monoester Hydrolases
    Phosphotransferases
    Genes
    Introns
    Messenger RNA
    Protein Isoforms
    Aptitude
    COS Cells
    Mitogen-Activated Protein Kinase 1
    Urinary Bladder Neoplasms
    Ovarian Neoplasms
    Exons
    Fibroblasts
    Breast Neoplasms
    Cell Line
    Skin
    Serum
    Proteins

    Keywords

    • Animals
    • COS Cells
    • Calcium-Calmodulin-Dependent Protein Kinases
    • Cytosol
    • Dual Specificity Phosphatase 6
    • Enzyme Activation
    • Fibroblasts
    • Fluorescent Antibody Technique
    • Gene Expression Regulation, Enzymologic
    • Growth Substances
    • Humans
    • JNK Mitogen-Activated Protein Kinases
    • Kinetics
    • Mitogen-Activated Protein Kinase 1
    • Mitogen-Activated Protein Kinases
    • Molecular Sequence Data
    • Phosphorylation
    • Protein Tyrosine Phosphatases
    • RNA, Messenger
    • Sequence Homology, Amino Acid
    • Signal Transduction
    • Skin
    • Stress, Physiological
    • Transfection

    Cite this

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    title = "Isolation of the human genes encoding the pyst1 and Pyst2 phosphatases: characterisation of Pyst2 as a cytosolic dual-specificity MAP kinase phosphatase and its catalytic activation by both MAP and SAP kinases",
    abstract = "We have isolated the human genes encoding the Pyst1 (MKP-3) and Pyst2 (MKP-X) MAP kinase phosphatases. Both genes consist of three exons interrupted by two introns and lack an intron which is conserved in all the other members of this gene family characterised to date. This reinforces the conclusion that Pyst1 and Pyst2 are members of a distinct and structurally homologous subfamily of dual-specificity (Thr/Tyr) MAP kinase phosphatases. We find that Pyst2 mRNA is constitutively expressed in a wide variety of human cell lines including those derived from ovarian, bladder and breast cancers. While there is no evidence for inducible expression of Pyst2 mRNA in human skin fibroblasts in response to cellular stress, Pyst2 mRNA levels are moderately increased in response to serum stimulation. Pyst2 protein is predominantly cytosolic when expressed in COS-1 cells. In common with Pyst1, Pyst2 shows substrate selectivity for the classical p42 (ERK2) isoform of MAP kinase both in vitro and in vivo, displaying much reduced activity towards stress activated MAP kinase isoforms such as JNK-1 and p38/RK. Pyst2 binds p42 MAP kinase in vivo and both MAP kinase binding and substrate selectivity correlate with the ability of different recombinant MAP and SAP kinases to cause catalytic activation of the Pyst2 phosphatase in vitro.",
    keywords = "Animals, COS Cells, Calcium-Calmodulin-Dependent Protein Kinases, Cytosol, Dual Specificity Phosphatase 6, Enzyme Activation, Fibroblasts, Fluorescent Antibody Technique, Gene Expression Regulation, Enzymologic, Growth Substances, Humans, JNK Mitogen-Activated Protein Kinases, Kinetics, Mitogen-Activated Protein Kinase 1, Mitogen-Activated Protein Kinases, Molecular Sequence Data, Phosphorylation, Protein Tyrosine Phosphatases, RNA, Messenger, Sequence Homology, Amino Acid, Signal Transduction, Skin, Stress, Physiological, Transfection",
    author = "S Dowd and Sneddon, {A A} and Keyse, {S M}",
    year = "1998",
    language = "English",
    volume = "111 ( Pt 22)",
    pages = "3389--99",
    journal = "Journal of Cell Science",
    issn = "0021-9533",
    publisher = "Company of Biologists Ltd",

    }

    TY - JOUR

    T1 - Isolation of the human genes encoding the pyst1 and Pyst2 phosphatases: characterisation of Pyst2 as a cytosolic dual-specificity MAP kinase phosphatase and its catalytic activation by both MAP and SAP kinases

    AU - Dowd, S

    AU - Sneddon, A A

    AU - Keyse, S M

    PY - 1998

    Y1 - 1998

    N2 - We have isolated the human genes encoding the Pyst1 (MKP-3) and Pyst2 (MKP-X) MAP kinase phosphatases. Both genes consist of three exons interrupted by two introns and lack an intron which is conserved in all the other members of this gene family characterised to date. This reinforces the conclusion that Pyst1 and Pyst2 are members of a distinct and structurally homologous subfamily of dual-specificity (Thr/Tyr) MAP kinase phosphatases. We find that Pyst2 mRNA is constitutively expressed in a wide variety of human cell lines including those derived from ovarian, bladder and breast cancers. While there is no evidence for inducible expression of Pyst2 mRNA in human skin fibroblasts in response to cellular stress, Pyst2 mRNA levels are moderately increased in response to serum stimulation. Pyst2 protein is predominantly cytosolic when expressed in COS-1 cells. In common with Pyst1, Pyst2 shows substrate selectivity for the classical p42 (ERK2) isoform of MAP kinase both in vitro and in vivo, displaying much reduced activity towards stress activated MAP kinase isoforms such as JNK-1 and p38/RK. Pyst2 binds p42 MAP kinase in vivo and both MAP kinase binding and substrate selectivity correlate with the ability of different recombinant MAP and SAP kinases to cause catalytic activation of the Pyst2 phosphatase in vitro.

    AB - We have isolated the human genes encoding the Pyst1 (MKP-3) and Pyst2 (MKP-X) MAP kinase phosphatases. Both genes consist of three exons interrupted by two introns and lack an intron which is conserved in all the other members of this gene family characterised to date. This reinforces the conclusion that Pyst1 and Pyst2 are members of a distinct and structurally homologous subfamily of dual-specificity (Thr/Tyr) MAP kinase phosphatases. We find that Pyst2 mRNA is constitutively expressed in a wide variety of human cell lines including those derived from ovarian, bladder and breast cancers. While there is no evidence for inducible expression of Pyst2 mRNA in human skin fibroblasts in response to cellular stress, Pyst2 mRNA levels are moderately increased in response to serum stimulation. Pyst2 protein is predominantly cytosolic when expressed in COS-1 cells. In common with Pyst1, Pyst2 shows substrate selectivity for the classical p42 (ERK2) isoform of MAP kinase both in vitro and in vivo, displaying much reduced activity towards stress activated MAP kinase isoforms such as JNK-1 and p38/RK. Pyst2 binds p42 MAP kinase in vivo and both MAP kinase binding and substrate selectivity correlate with the ability of different recombinant MAP and SAP kinases to cause catalytic activation of the Pyst2 phosphatase in vitro.

    KW - Animals

    KW - COS Cells

    KW - Calcium-Calmodulin-Dependent Protein Kinases

    KW - Cytosol

    KW - Dual Specificity Phosphatase 6

    KW - Enzyme Activation

    KW - Fibroblasts

    KW - Fluorescent Antibody Technique

    KW - Gene Expression Regulation, Enzymologic

    KW - Growth Substances

    KW - Humans

    KW - JNK Mitogen-Activated Protein Kinases

    KW - Kinetics

    KW - Mitogen-Activated Protein Kinase 1

    KW - Mitogen-Activated Protein Kinases

    KW - Molecular Sequence Data

    KW - Phosphorylation

    KW - Protein Tyrosine Phosphatases

    KW - RNA, Messenger

    KW - Sequence Homology, Amino Acid

    KW - Signal Transduction

    KW - Skin

    KW - Stress, Physiological

    KW - Transfection

    M3 - Article

    VL - 111 ( Pt 22)

    SP - 3389

    EP - 3399

    JO - Journal of Cell Science

    JF - Journal of Cell Science

    SN - 0021-9533

    ER -