Akt binds to and phosphorylates phospholipase C-gamma1 in response to epidermal growth factor.

Both phospholipase (PL) C-gamma1 and Akt (protein kinase B; PKB) are signaling proteins that play significant roles in the intracellular signaling mechanism used by receptor tyrosine kinases, including epidermal growth factor (EGF) receptor (EGFR). EGFR activates PLC-gamma1 directly and activates Akt indirectly through phosphatidylinositol 3-kinase (PI3K). Many studies have shown that the PLC-gamma1 pathway and PI3K-Akt pathway interact with each other. However, it is not known whether PLC-gamma1 binds to Akt directly. In this communication, we identified a novel interaction between PLC-gamma1 and Akt. We demonstrated that the interaction is mediated by the binding of PLC-gamma1 Src homology (SH) 3 domain to Akt proline-rich motifs. We also provide a novel model to depict how the interaction between PLC-gamma1 SH3 domain and Akt proline-rich motifs is dependent on EGF stimulation. In this model, phosphorylation of PLC-gamma1 Y783 by EGF causes the conformational change of PLC-gamma1 to allow the interaction of its SH3 domain with Akt proline-rich motifs. Furthermore, we showed that the interaction between PLC-gamma1 and Akt resulted in the phosphorylation of PLC-gamma1 S1248 by Akt. Finally, we showed that the interaction between PLC-gamma1 and Akt enhanced EGF-stimulated cell motility.

[1]  S. Rhee,et al.  Intramolecular interaction between phosphorylated tyrosine-783 and the C-terminal Src homology 2 domain activates phospholipase C-gamma1. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[2]  G. Carpenter,et al.  Selective phospholipase C activation , 1991, BioEssays : news and reviews in molecular, cellular and developmental biology.

[3]  P. Cohen,et al.  Role of Translocation in the Activation and Function of Protein Kinase B* , 1997, The Journal of Biological Chemistry.

[4]  A. Ullrich,et al.  SH2 domains prevent tyrosine dephosphorylation of the EGF receptor: identification of Tyr992 as the high‐affinity binding site for SH2 domains of phospholipase C gamma. , 1992, The EMBO journal.

[5]  S. Rhee,et al.  Phosphorylation of phospholipase C-gamma by cAMP-dependent protein kinase. , 1989, The Journal of biological chemistry.

[6]  B. Zetter,et al.  Regulation of chemotaxis by the platelet-derived growth factor receptor-β , 1994, Nature.

[7]  R. Jorissen,et al.  Epidermal growth factor receptor , 2003 .

[8]  A. Wells,et al.  Epidermal growth factor receptor‐mediated motility in fibroblasts , 1998, Microscopy research and technique.

[9]  Frederick Y. Wu,et al.  Phospholipase Cγ1 is a physiological guanine nucleotide exchange factor for the nuclear GTPase PIKE , 2002, Nature.

[10]  Michael J. Fry,et al.  Phosphatidylinositol-3-OH kinase direct target of Ras , 1994, Nature.

[11]  T. K. Yeung,et al.  Endocytosis deficiency of epidermal growth factor (EGF) receptor-ErbB2 heterodimers in response to EGF stimulation. , 1999, Molecular biology of the cell.

[12]  X. Chen,et al.  Regulation of intracellular trafficking of the EGF receptor by Rab5 in the absence of phosphatidylinositol 3‐kinase activity , 2001, EMBO reports.

[13]  M. Moran,et al.  Phospholipase C-gamma1: a phospholipase and guanine nucleotide exchange factor. , 2002, Molecular interventions.

[14]  M. David,et al.  Distinct Mechanisms Determine the Patterns of Differential Activation of H-Ras, N-Ras, K-Ras 4B, and M-Ras by Receptors for Growth Factors or Antigen , 2004, Molecular and Cellular Biology.

[15]  M. Beaven,et al.  Certain inhibitors of protein serine/threonine kinases also inhibit tyrosine phosphorylation of phospholipase C gamma 1 and other proteins and reveal distinct roles for tyrosine kinase(s) and protein kinase C in stimulated, rat basophilic RBL-2H3 cells. , 1992, Journal of immunology.

[16]  G. Carpenter,et al.  Phospholipase C-gamma1: regulation of enzyme function and role in growth factor-dependent signal transduction. , 1997, Cytokine & growth factor reviews.

[17]  M. Moran,et al.  Binding of SH2 domains of phospholipase C gamma 1, GAP, and Src to activated growth factor receptors. , 1990, Science.

[18]  A. Takayanagi,et al.  A novel gene delivery system using EGF receptor‐mediated endocytosis , 1994, FEBS letters.

[19]  S. R. Datta,et al.  Akt Phosphorylation of BAD Couples Survival Signals to the Cell-Intrinsic Death Machinery , 1997, Cell.

[20]  Kyong-Tai Kim,et al.  The Direct Interaction of Phospholipase C-γ1 with Phospholipase D2 Is Important for Epidermal Growth Factor Signaling* , 2003, The Journal of Biological Chemistry.

[21]  J. Grandis,et al.  Epidermal Growth Factor Receptor-stimulated Activation of Phospholipase Cγ-1 Promotes Invasion of Head and Neck Squamous Cell Carcinoma , 2003 .

[22]  U. Smith,et al.  Impaired phosphorylation and insulin-stimulated translocation to the plasma membrane of protein kinase B/Akt in adipocytes from Type II diabetic subjects , 2000, Diabetologia.

[23]  N. Rahimi,et al.  Recruitment and Activation of Phospholipase Cγ1 by Vascular Endothelial Growth Factor Receptor-2 Are Required for Tubulogenesis and Differentiation of Endothelial Cells* , 2003, The Journal of Biological Chemistry.

[24]  L. Peso,et al.  Interleukin-3-induced phosphorylation of BAD through the protein kinase Akt. , 1997, Science.

[25]  S. Snyder,et al.  Phospholipase C-γ1 is a guanine nucleotide exchange factor for dynamin-1 and enhances dynamin-1-dependent epidermal growth factor receptor endocytosis , 2004, Journal of Cell Science.

[26]  Simon C Watkins,et al.  Directional motility induced by epidermal growth factor requires Cdc42. , 2003, Experimental cell research.

[27]  R. Dickson,et al.  Calmodulin-mediated Activation of Akt Regulates Survival of c-Myc-overexpressing Mouse Mammary Carcinoma Cells* , 2004, Journal of Biological Chemistry.

[28]  J. Grandis,et al.  Phospholipase C-γ1 in tumor progression , 2003, Clinical & Experimental Metastasis.

[29]  A. Zilberstein,et al.  EGF induces tyrosine phosphorylation of phospholipase C-II: A potential mechanism for EGF receptor signaling , 1989, Cell.

[30]  E. Vigorito,et al.  PLCγ2 regulates Bcl‐2 levels and is required for survival rather than differentiation of marginal zone and follicular B cells , 2004, European journal of immunology.

[31]  Alan Wells,et al.  Growth Factor-Induced Cell Motility in Tumor Invasion , 2002, Acta oncologica.

[32]  L. Cantley,et al.  An 11-Amino Acid Sequence from c-met Initiates Epithelial Chemotaxis via Phosphatidylinositol 3-Kinase and Phospholipase C (*) , 1996, The Journal of Biological Chemistry.

[33]  A. Ullrich,et al.  Effect of phospholipase C-gamma overexpression on PDGF-induced second messengers and mitogenesis. , 1990, Science.

[34]  S. Kanner,et al.  Regulated association between the tyrosine kinase Emt/Itk/Tsk and phospholipase-C gamma 1 in human T lymphocytes. , 1999, Journal of immunology.

[35]  U. Hellman,et al.  Identification of two C‐terminal autophosphorylation sites in the PDGF beta‐receptor: involvement in the interaction with phospholipase C‐gamma. , 1992, The EMBO journal.

[36]  Zhixiang Wang,et al.  Regulation of EGF‐Induced Phospholipase C‐γ1 Translocation and Activation by its SH2 and PH Domains , 2003, Traffic.

[37]  G. Carpenter,et al.  Antiphosphotyrosine recovery of phospholipase C activity after EGF treatment of A-431 cells. , 1988, Science.

[38]  S. Ryu,et al.  Direct interaction of SOS1 Ras exchange protein with the SH3 domain of phospholipase C-gamma1. , 2000, Biochemistry.

[39]  Peter J. Cullen,et al.  Phospholipase Cγ activates Ras on the Golgi apparatus by means of RasGRP1 , 2003, Nature.

[40]  R. Firtel,et al.  The molecular genetics of chemotaxis: sensing and responding to chemoattractant gradients , 2000, BioEssays : news and reviews in molecular, cellular and developmental biology.

[41]  S. Snyder,et al.  Phospholipase C-γ Is Required for Agonist-Induced Ca2+ Entry , 2002, Cell.

[42]  A. Zilberstein,et al.  PDGF stimulation of inositol phospholipid hydrolysis requires PLC-γ1 phosphorylation on tyrosine residues 783 and 1254 , 1991, Cell.

[43]  P. Comoglio,et al.  Sustained recruitment of phospholipase C-γ to Gab1 is required for HGF-induced branching tubulogenesis , 2000, Oncogene.

[44]  P. Cohen,et al.  Mechanism of activation of protein kinase B by insulin and IGF‐1. , 1996, The EMBO journal.

[45]  G. Carpenter,et al.  Phospholipase C-gamma as a signal-transducing element. , 1999, Experimental cell research.

[46]  T. Hunter,et al.  Phospholipase C-γ is a substrate for the PDGF and EGF receptor protein-tyrosine kinases in vivo and in vitro , 1989, Cell.

[47]  Michael F. Moran,et al.  Requirement for Phospholipase C-γ1 Enzymatic Activity in Growth Factor-Induced Mitogenesis , 1998, Molecular and Cellular Biology.

[48]  B. Hemmings,et al.  Structure, regulation and function of PKB/AKT--a major therapeutic target. , 2004, Biochimica et biophysica acta.

[49]  T. Pawson,et al.  Activated Src tyrosine kinase phosphorylates Tyr-457 of bovine GTPase-activating protein (GAP) in vitro and the corresponding residue of rat GAP in vivo. , 1992, The Journal of biological chemistry.