ZNRF2 is released from membranes by growth factors and with ZNRF1 regulates the Na+/K+ATPase

    Research output: Contribution to journalArticle

    10 Citations (Scopus)

    Abstract

    Here, we describe a phosphorylation-based reverse myristoyl switch for mammalian ZNRF2, and show that this E3 ubiquitin ligase and its sister protein ZNRF1 regulate the Na+/K+ pump (Na+/K+ATPase). N-myristoylation localizes ZNRF1 and ZNRF2 to intracellular membranes and enhances their activity. However, when ZNRF2 is phosphorylated in response to agonists including insulin and growth factors, it binds to 14-3-3 and is released into the cytosol. On membranes, ZNRF1 and ZNRF2 interact with the Na+/K+ATPase a1 subunit via their UBZ domains, while their RING domains interact with E2 proteins, predominantly Ubc13 that, together with Uev1a, mediates formation of Lys63-ubiquitin linkages. ZNRF1 and ZNRF2 can ubiquitylate the cytoplasmic loop encompassing the nucleotide-binding and phosphorylation regions of the Na+/K+ATPase a1 subunit. Ouabain, a Na+/K+ATPase inhibitor and therapeutic cardiac glycoside, decreases ZNRF1 protein levels, whereas knockdown of ZNRF2 inhibits the ouabain-induced decrease of cell surface and total Na+/K+ATPase a1 levels. Thus, ZNRF1 and ZNRF2 are new players in regulation of the ubiquitous Na+/K+ATPase that is tuned to changing demands in many physiological contexts.

    Original languageEnglish
    Pages (from-to)4662-4675
    Number of pages14
    JournalJournal of Cell Science
    Volume125
    Issue number19
    DOIs
    Publication statusPublished - 1 Oct 2012

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    Intercellular Signaling Peptides and Proteins
    Membranes
    Ouabain
    Phosphorylation
    Cardiac Glycosides
    Intracellular Membranes
    Proteins
    Ubiquitin-Protein Ligases
    Ubiquitin
    Cytosol
    Nucleotides
    sodium-translocating ATPase
    Insulin
    Therapeutics

    Keywords

    • 14-3-3 Proteins
    • Amino Acid Sequence
    • Animals
    • Carrier Proteins
    • Cytoplasm
    • Gene Knockdown Techniques
    • HEK293 Cells
    • Humans
    • Intercellular Signaling Peptides and Proteins
    • Intracellular Membranes
    • Isotope Labeling
    • Mice
    • Molecular Sequence Data
    • Mutation
    • Ouabain
    • Phosphorylation
    • Phosphoserine
    • Protein Binding
    • Protein Structure, Tertiary
    • Protein Subunits
    • Protein Transport
    • Sodium-Potassium-Exchanging ATPase
    • Ubiquitin-Conjugating Enzymes
    • Ubiquitin-Protein Ligases
    • Ubiquitination

    Cite this

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    title = "ZNRF2 is released from membranes by growth factors and with ZNRF1 regulates the Na+/K+ATPase",
    abstract = "Here, we describe a phosphorylation-based reverse myristoyl switch for mammalian ZNRF2, and show that this E3 ubiquitin ligase and its sister protein ZNRF1 regulate the Na+/K+ pump (Na+/K+ATPase). N-myristoylation localizes ZNRF1 and ZNRF2 to intracellular membranes and enhances their activity. However, when ZNRF2 is phosphorylated in response to agonists including insulin and growth factors, it binds to 14-3-3 and is released into the cytosol. On membranes, ZNRF1 and ZNRF2 interact with the Na+/K+ATPase a1 subunit via their UBZ domains, while their RING domains interact with E2 proteins, predominantly Ubc13 that, together with Uev1a, mediates formation of Lys63-ubiquitin linkages. ZNRF1 and ZNRF2 can ubiquitylate the cytoplasmic loop encompassing the nucleotide-binding and phosphorylation regions of the Na+/K+ATPase a1 subunit. Ouabain, a Na+/K+ATPase inhibitor and therapeutic cardiac glycoside, decreases ZNRF1 protein levels, whereas knockdown of ZNRF2 inhibits the ouabain-induced decrease of cell surface and total Na+/K+ATPase a1 levels. Thus, ZNRF1 and ZNRF2 are new players in regulation of the ubiquitous Na+/K+ATPase that is tuned to changing demands in many physiological contexts.",
    keywords = "14-3-3 Proteins, Amino Acid Sequence, Animals, Carrier Proteins, Cytoplasm, Gene Knockdown Techniques, HEK293 Cells, Humans, Intercellular Signaling Peptides and Proteins, Intracellular Membranes, Isotope Labeling, Mice, Molecular Sequence Data, Mutation, Ouabain, Phosphorylation, Phosphoserine, Protein Binding, Protein Structure, Tertiary, Protein Subunits, Protein Transport, Sodium-Potassium-Exchanging ATPase, Ubiquitin-Conjugating Enzymes, Ubiquitin-Protein Ligases, Ubiquitination",
    author = "Gerta Hoxhaj and Ayaz Najafov and Rachel Toth and Campbell, {David G.} and Prescott, {Alan R.} and Carol MacKintosh",
    year = "2012",
    month = "10",
    day = "1",
    doi = "10.1242/jcs.110296",
    language = "English",
    volume = "125",
    pages = "4662--4675",
    journal = "Journal of Cell Science",
    issn = "0021-9533",
    publisher = "Company of Biologists",
    number = "19",

    }

    ZNRF2 is released from membranes by growth factors and with ZNRF1 regulates the Na+/K+ATPase. / Hoxhaj, Gerta; Najafov, Ayaz; Toth, Rachel; Campbell, David G.; Prescott, Alan R.; MacKintosh, Carol.

    In: Journal of Cell Science, Vol. 125, No. 19, 01.10.2012, p. 4662-4675.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - ZNRF2 is released from membranes by growth factors and with ZNRF1 regulates the Na+/K+ATPase

    AU - Hoxhaj, Gerta

    AU - Najafov, Ayaz

    AU - Toth, Rachel

    AU - Campbell, David G.

    AU - Prescott, Alan R.

    AU - MacKintosh, Carol

    PY - 2012/10/1

    Y1 - 2012/10/1

    N2 - Here, we describe a phosphorylation-based reverse myristoyl switch for mammalian ZNRF2, and show that this E3 ubiquitin ligase and its sister protein ZNRF1 regulate the Na+/K+ pump (Na+/K+ATPase). N-myristoylation localizes ZNRF1 and ZNRF2 to intracellular membranes and enhances their activity. However, when ZNRF2 is phosphorylated in response to agonists including insulin and growth factors, it binds to 14-3-3 and is released into the cytosol. On membranes, ZNRF1 and ZNRF2 interact with the Na+/K+ATPase a1 subunit via their UBZ domains, while their RING domains interact with E2 proteins, predominantly Ubc13 that, together with Uev1a, mediates formation of Lys63-ubiquitin linkages. ZNRF1 and ZNRF2 can ubiquitylate the cytoplasmic loop encompassing the nucleotide-binding and phosphorylation regions of the Na+/K+ATPase a1 subunit. Ouabain, a Na+/K+ATPase inhibitor and therapeutic cardiac glycoside, decreases ZNRF1 protein levels, whereas knockdown of ZNRF2 inhibits the ouabain-induced decrease of cell surface and total Na+/K+ATPase a1 levels. Thus, ZNRF1 and ZNRF2 are new players in regulation of the ubiquitous Na+/K+ATPase that is tuned to changing demands in many physiological contexts.

    AB - Here, we describe a phosphorylation-based reverse myristoyl switch for mammalian ZNRF2, and show that this E3 ubiquitin ligase and its sister protein ZNRF1 regulate the Na+/K+ pump (Na+/K+ATPase). N-myristoylation localizes ZNRF1 and ZNRF2 to intracellular membranes and enhances their activity. However, when ZNRF2 is phosphorylated in response to agonists including insulin and growth factors, it binds to 14-3-3 and is released into the cytosol. On membranes, ZNRF1 and ZNRF2 interact with the Na+/K+ATPase a1 subunit via their UBZ domains, while their RING domains interact with E2 proteins, predominantly Ubc13 that, together with Uev1a, mediates formation of Lys63-ubiquitin linkages. ZNRF1 and ZNRF2 can ubiquitylate the cytoplasmic loop encompassing the nucleotide-binding and phosphorylation regions of the Na+/K+ATPase a1 subunit. Ouabain, a Na+/K+ATPase inhibitor and therapeutic cardiac glycoside, decreases ZNRF1 protein levels, whereas knockdown of ZNRF2 inhibits the ouabain-induced decrease of cell surface and total Na+/K+ATPase a1 levels. Thus, ZNRF1 and ZNRF2 are new players in regulation of the ubiquitous Na+/K+ATPase that is tuned to changing demands in many physiological contexts.

    KW - 14-3-3 Proteins

    KW - Amino Acid Sequence

    KW - Animals

    KW - Carrier Proteins

    KW - Cytoplasm

    KW - Gene Knockdown Techniques

    KW - HEK293 Cells

    KW - Humans

    KW - Intercellular Signaling Peptides and Proteins

    KW - Intracellular Membranes

    KW - Isotope Labeling

    KW - Mice

    KW - Molecular Sequence Data

    KW - Mutation

    KW - Ouabain

    KW - Phosphorylation

    KW - Phosphoserine

    KW - Protein Binding

    KW - Protein Structure, Tertiary

    KW - Protein Subunits

    KW - Protein Transport

    KW - Sodium-Potassium-Exchanging ATPase

    KW - Ubiquitin-Conjugating Enzymes

    KW - Ubiquitin-Protein Ligases

    KW - Ubiquitination

    U2 - 10.1242/jcs.110296

    DO - 10.1242/jcs.110296

    M3 - Article

    VL - 125

    SP - 4662

    EP - 4675

    JO - Journal of Cell Science

    JF - Journal of Cell Science

    SN - 0021-9533

    IS - 19

    ER -