Identification of RhoGAP22 as an Akt-dependent regulator of cell motility in response to insulin

Alexander F. Rowland (Lead / Corresponding author), Mark Larance (Lead / Corresponding author), William E. Hughes, David E. James (Lead / Corresponding author)

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    11 Citations (Scopus)

    Abstract

    Insulin exerts many of its metabolic actions via the canonical phosphatidylinositide 3 kinase (PI3K)/Akt pathway, leading to phosphorylation and 14-3-3 binding of key metabolic targets. We previously identified a GTPase-activating protein (GAP) for Rac1 called RhoGAP22 as an insulin-responsive 14-3-3 binding protein. Insulin increased 14-3-3 binding to RhoGAP22 fourfold, and this effect was PI3K dependent. We identified two insulin-responsive 14-3-3 binding sites (pSer(16) and pSer(395)) within RhoGAP22, and mutagenesis studies revealed a complex interplay between the phosphorylation at these two sites. Mutating Ser(16) to alanine blocked 14-3-3 binding to RhoGAP22 in vivo, and phosphorylation at Ser(16) was mediated by the kinase Akt. Overexpression of a mutant RhoGAP22 that was unable to bind 14-3-3 reduced cell motility in NIH-3T3 fibroblasts, and this effect was dependent on a functional GAP domain. Mutation of the catalytic arginine of the GAP domain of RhoGAP22 potentiated growth factor-stimulated Rac1 GTP loading. We propose that insulin and possibly growth factors such as platelet-derived growth factor may play a novel role in regulating cell migration and motility via the Akt-dependent phosphorylation of RhoGAP22, leading to modulation of Rac1 activity.
    Original languageEnglish
    Pages (from-to)4789-4800
    Number of pages12
    JournalMolecular and Cellular Biology
    Volume31
    Issue number23
    DOIs
    Publication statusPublished - Dec 2011

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    Keywords

    • Cell Movement
    • Animals
    • 14-3-3 Proteins
    • GTPase-Activating Proteins
    • Humans
    • Catalytic Domain
    • Proto-Oncogene Proteins c-akt
    • Mice
    • Amino Acid Sequence
    • Protein Binding
    • Insulin
    • Binding Sites
    • Rats
    • Gene Knockdown Techniques
    • Phosphorylation
    • Conserved Sequence
    • Molecular Sequence Data
    • RNA Interference
    • Protein Structure, Tertiary
    • Amino Acid Substitution
    • Cell Line
    • Cricetinae

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