Phosphoregulation of Arp2/3-dependent actin assembly during receptor-mediated endocytosis

Jiro Toshima, Junko Y. Toshima, Adam C. Martin, David G. Drubin

    Research output: Contribution to journalArticle

    78 Citations (Scopus)

    Abstract

    In both yeast and mammals, endocytic internalization is accompanied by a transient burst of actin polymerization. The yeast protein kinases Prk1p and Ark1p, which are related to the mammalian proteins GAK and AAK1, are key regulators of this process. However, the molecular mechanism(s) by which they regulate actin assembly at endocytic sites have not yet been determined. The Eps15-like yeast protein Pan1p is a Prk1p substrate that is essential for endocytic internalization and for proper actin organization. Pan1p is an Arp2/3 activator and here we show that this activity is dependent on F-actin binding. Mutation of all 15 Prk1p-targeted threonines in Pan1p to alanines mimicked the ark1Δ prk1Δ phenotype, demonstrating that Pan1p is a key Prk1p target in vivo. Moreover, phosphorylation by Prk1p inhibited the ability of Pan1p to bind to F-actin and to activate the Arp2/3 complex, thereby identifying the endocytic phosphoregulation mechanism of Prk1p. We conclude that Prk1p phosphorylation of Pan1p shuts off Arp2/3-mediated actin polymerization on endocytic vesicles, allowing them to fuse with endosomes.

    Original languageEnglish
    Pages (from-to)246-254
    Number of pages9
    JournalNature Cell Biology
    Volume7
    Issue number3
    DOIs
    Publication statusPublished - 2005 Mar

    Fingerprint

    Endocytosis
    Actins
    Fungal Proteins
    Polymerization
    Actin-Related Protein 2-3 Complex
    Phosphorylation
    Transport Vesicles
    Endosomes
    Threonine
    Alanine
    Protein Kinases
    Mammals
    Yeasts
    Phenotype
    Mutation
    Proteins

    ASJC Scopus subject areas

    • Cell Biology

    Cite this

    Phosphoregulation of Arp2/3-dependent actin assembly during receptor-mediated endocytosis. / Toshima, Jiro; Toshima, Junko Y.; Martin, Adam C.; Drubin, David G.

    In: Nature Cell Biology, Vol. 7, No. 3, 03.2005, p. 246-254.

    Research output: Contribution to journalArticle

    Toshima, Jiro ; Toshima, Junko Y. ; Martin, Adam C. ; Drubin, David G. / Phosphoregulation of Arp2/3-dependent actin assembly during receptor-mediated endocytosis. In: Nature Cell Biology. 2005 ; Vol. 7, No. 3. pp. 246-254.
    @article{b9ada4ad0add426da56a7533de76c1b3,
    title = "Phosphoregulation of Arp2/3-dependent actin assembly during receptor-mediated endocytosis",
    abstract = "In both yeast and mammals, endocytic internalization is accompanied by a transient burst of actin polymerization. The yeast protein kinases Prk1p and Ark1p, which are related to the mammalian proteins GAK and AAK1, are key regulators of this process. However, the molecular mechanism(s) by which they regulate actin assembly at endocytic sites have not yet been determined. The Eps15-like yeast protein Pan1p is a Prk1p substrate that is essential for endocytic internalization and for proper actin organization. Pan1p is an Arp2/3 activator and here we show that this activity is dependent on F-actin binding. Mutation of all 15 Prk1p-targeted threonines in Pan1p to alanines mimicked the ark1Δ prk1Δ phenotype, demonstrating that Pan1p is a key Prk1p target in vivo. Moreover, phosphorylation by Prk1p inhibited the ability of Pan1p to bind to F-actin and to activate the Arp2/3 complex, thereby identifying the endocytic phosphoregulation mechanism of Prk1p. We conclude that Prk1p phosphorylation of Pan1p shuts off Arp2/3-mediated actin polymerization on endocytic vesicles, allowing them to fuse with endosomes.",
    author = "Jiro Toshima and Toshima, {Junko Y.} and Martin, {Adam C.} and Drubin, {David G.}",
    year = "2005",
    month = "3",
    doi = "10.1038/ncb1229",
    language = "English",
    volume = "7",
    pages = "246--254",
    journal = "Nature Cell Biology",
    issn = "1465-7392",
    publisher = "Nature Publishing Group",
    number = "3",

    }

    TY - JOUR

    T1 - Phosphoregulation of Arp2/3-dependent actin assembly during receptor-mediated endocytosis

    AU - Toshima, Jiro

    AU - Toshima, Junko Y.

    AU - Martin, Adam C.

    AU - Drubin, David G.

    PY - 2005/3

    Y1 - 2005/3

    N2 - In both yeast and mammals, endocytic internalization is accompanied by a transient burst of actin polymerization. The yeast protein kinases Prk1p and Ark1p, which are related to the mammalian proteins GAK and AAK1, are key regulators of this process. However, the molecular mechanism(s) by which they regulate actin assembly at endocytic sites have not yet been determined. The Eps15-like yeast protein Pan1p is a Prk1p substrate that is essential for endocytic internalization and for proper actin organization. Pan1p is an Arp2/3 activator and here we show that this activity is dependent on F-actin binding. Mutation of all 15 Prk1p-targeted threonines in Pan1p to alanines mimicked the ark1Δ prk1Δ phenotype, demonstrating that Pan1p is a key Prk1p target in vivo. Moreover, phosphorylation by Prk1p inhibited the ability of Pan1p to bind to F-actin and to activate the Arp2/3 complex, thereby identifying the endocytic phosphoregulation mechanism of Prk1p. We conclude that Prk1p phosphorylation of Pan1p shuts off Arp2/3-mediated actin polymerization on endocytic vesicles, allowing them to fuse with endosomes.

    AB - In both yeast and mammals, endocytic internalization is accompanied by a transient burst of actin polymerization. The yeast protein kinases Prk1p and Ark1p, which are related to the mammalian proteins GAK and AAK1, are key regulators of this process. However, the molecular mechanism(s) by which they regulate actin assembly at endocytic sites have not yet been determined. The Eps15-like yeast protein Pan1p is a Prk1p substrate that is essential for endocytic internalization and for proper actin organization. Pan1p is an Arp2/3 activator and here we show that this activity is dependent on F-actin binding. Mutation of all 15 Prk1p-targeted threonines in Pan1p to alanines mimicked the ark1Δ prk1Δ phenotype, demonstrating that Pan1p is a key Prk1p target in vivo. Moreover, phosphorylation by Prk1p inhibited the ability of Pan1p to bind to F-actin and to activate the Arp2/3 complex, thereby identifying the endocytic phosphoregulation mechanism of Prk1p. We conclude that Prk1p phosphorylation of Pan1p shuts off Arp2/3-mediated actin polymerization on endocytic vesicles, allowing them to fuse with endosomes.

    UR - http://www.scopus.com/inward/record.url?scp=14744304957&partnerID=8YFLogxK

    UR - http://www.scopus.com/inward/citedby.url?scp=14744304957&partnerID=8YFLogxK

    U2 - 10.1038/ncb1229

    DO - 10.1038/ncb1229

    M3 - Article

    C2 - 15711538

    AN - SCOPUS:14744304957

    VL - 7

    SP - 246

    EP - 254

    JO - Nature Cell Biology

    JF - Nature Cell Biology

    SN - 1465-7392

    IS - 3

    ER -