Phosphatidylinositol 4,5‐bisphosphate stimulates phosphorylation of the adaptor protein Shc by c‐Src

Sato, Kenichi; Yamamoto, Hideki; Otsuki, Tetsuji; Aoto, Mamoru; Tokmakov, Alexander A.; Hayashi, Fumio; Fukami, Yasuo

doi:10.1016/s0014-5793(97)00539-5

Cited by 22 publications

(15 citation statements)

References 17 publications

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“…Others have shown a role for Shc Tyr239-Tyr240 and Shc Tyr317 phosphorylation in Grb2 binding and Ras-MAP kinase pathway activation (26,32). In addition, phosphorylation of Tyr239-Tyr240 in response to cytokines has been implicated in a Ras-independent pathway leading to c-Myc (8), and phosphorylation of these two residues by Src appears to be stimulated in some way by PIP2 binding to Shc (27,28). To explore the potential role of Shc molecules phosphorylated at each of these sites, we tested the effects of wild-type and mutant Shc proteins on PDGF-stimulated DNA synthesis, using plasmid DNA microinjection.…”

Section: Resultsmentioning

confidence: 99%

“…A Shc ⌬PTB domain deletion mutant also blocked PDGF-stimulated DNA synthesis. Others have shown that the Shc PTB domain binding to PIP2 may stimulate Src phosphorylation of Tyr239-Tyr240 through an as-yet-unknown mechanism-perhaps by causing colocalization of Src and Shc following growth factor receptor activation of phosphatidylinositol 3 kinase (27,28). Both inhibitory Shc mutants (2Y2F-R401K and ⌬PTB) could be rescued by Myc coexpression but not by Fos, suggesting that these Shc domains are required in the Src pathway.…”

Section: Discussionmentioning

confidence: 99%

“…Tyrosine phosphorylation at both Tyr239-Tyr240 and Tyr317 has been implicated in Grb2 binding and mitogen-activated protein (MAP) kinase pathway activation (8,20,26,32). Furthermore, activated versions of Src are capable of stimulating the Ras-MAP kinase pathway (34), and the principal mechanism is believed to involve the phosphorylation of Shc by Src (17,28,32), followed by Grb2 and Sos recruitment. The activation of the Ras-MAP kinase pathway by G protein-coupled receptors also appears to be Src dependent and mediated by Shc phosphorylation (17,18).…”

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confidence: 99%

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SU6656, a Selective Src Family Kinase Inhibitor, Used To Probe Growth Factor Signaling

Blake¹,

Broome²,

Liu³

et al. 2000

Molecular and Cellular Biology

548

463

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The use of small-molecule inhibitors to study molecular components of cellular signal transduction pathways provides a means of analysis complementary to currently used techniques, such as antisense, dominantnegative (interfering) mutants and constitutively activated mutants. We have identified and characterized a small-molecule inhibitor, SU6656, which exhibits selectivity for Src and other members of the Src family. A related inhibitor, SU6657, inhibits many kinases, including Src and the platelet-derived growth factor (PDGF) receptor. The use of SU6656 confirmed our previous findings that Src family kinases are required for both Myc induction and DNA synthesis in response to PDGF stimulation of NIH 3T3 fibroblasts. By comparing PDGFstimulated tyrosine phosphorylation events in untreated and SU6656-treated cells, we found that some substrates (for example, c-Cbl, and protein kinase C ␦) were Src family substrates whereas others (for example, phospholipase C-␥) were not. One protein, the adaptor Shc, was a substrate for both Src family kinases (on tyrosines 239 and 240) and a distinct tyrosine kinase (on tyrosine 317, which is perhaps phosphorylated by the PDGF receptor itself). Microinjection experiments demonstrated that a Shc molecule carrying mutations of tyrosines 239 and 240, in conjunction with an SH2 domain mutation, interfered with PDGF-stimulated DNA synthesis. Deletion of the phosphotyrosine-binding domain also inhibited synthesis. These inhibitions were overcome by heterologous expression of Myc, supporting the hypothesis that Shc functions in the Src pathway. SU6656 should prove a useful additional tool for further dissecting the role of Src kinases in this and other signal transduction pathways.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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SU6656, a Selective Src Family Kinase Inhibitor, Used To Probe Growth Factor Signaling

Blake¹,

Broome²,

Liu³

et al. 2000

Molecular and Cellular Biology

548

463

View full text Add to dashboard Cite

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“…Amounts of GST fusion proteins immobilized on Sepharose 4B beads were detected by Ponceau stain. Stars denote GST-fusion proteins of correct molecular weight (p52 and CH domain contained proteolytic breakdown products) (29). Data shown are representative of 2 experiments performed.…”

Section: Attenuated Activation Of C-src Kinase In Cells Deficient In mentioning

confidence: 99%

Plasma Membrane Phospholipid Scramblase 1 Promotes EGF-dependent Activation of c-Src through the Epidermal Growth Factor Receptor

Nanjundan

Sun²,

Zhao

et al. 2003

Journal of Biological Chemistry

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Phospholipid scramblase (PLSCR1) is a multiply palmitoylated, calcium-binding endofacial membrane protein proposed to mediate transbilayer movement of plasma membrane phospholipids. PLSCR1 is a component of membrane lipid rafts and has been shown to both physically and functionally interact with activated epidermal growth factor (EGF) receptors and other raft-associated cell surface receptors. Cell stimulation by EGF results in Tyr phosphorylation of PLSCR1, its association with both Shc and EGF receptors, and rapid cycling of PLSCR1 between plasma membrane and endosomal compartments. We now report evidence that upon EGF stimulation, PLSCR1 is phosphorylated by c-Src, within the tandem repeat sequence 68 VYNQPVYNQP 77. The in vivo interaction between PLSCR1 and Shc requires the Srcmediated phosphorylation on tyrosines 69 and 74. In in vitro pull down studies, phosphorylated PLSCR1 was found to bind directly to Shc through the phosphotyrosine binding domain. Consistent with the potential role of PLSCR1 in growth factor signaling pathways, granulocyte precursors derived from mice deficient in PLSCR1 show impaired proliferation and maturation under cytokine stimulation. Using PLSCR1؊/؊ embryonic fibroblasts and kidney epithelial cells, we now demonstrate that deletion of PLSCR1 from the plasma membrane reduces the activation of c-Src by EGF, implying that PLSCR1 normally facilitates receptor-dependent activation of this kinase. We propose that PLSCR1, through its interaction with Shc, promotes Src kinase activation through the EGF receptor.

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“…To obtain GFP-Shc expression constructs, cDNAs encoding mouse Shc isoforms of P46 (amino acids residues 46±469 of P52) and P52 (1±469 of P52), in the expression vector pGEX-2T (Pharmacia), were isolated as previously described (Sato et al 1997a). Each pGEX-2T/Shc construct was digested with EcoRI and the resultant digest containing each Shc gene was subcloned into the EcoRI site of pEGFP-C3 (Clontech laboratories, CA, USA).…”

Section: Plasmid Constructionsmentioning

confidence: 99%

Adaptor protein Shc undergoes translocation and mediates up‐regulation of the tyrosine kinase c‐Src in EGF‐stimulated A431 cells

Sato

Kimoto²,

Kakumoto³

et al. 2000

Genes to Cells

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Phosphatidylinositol 4,5‐bisphosphate stimulates phosphorylation of the adaptor protein Shc by c‐Src

Cited by 22 publications

References 17 publications

SU6656, a Selective Src Family Kinase Inhibitor, Used To Probe Growth Factor Signaling

SU6656, a Selective Src Family Kinase Inhibitor, Used To Probe Growth Factor Signaling

Plasma Membrane Phospholipid Scramblase 1 Promotes EGF-dependent Activation of c-Src through the Epidermal Growth Factor Receptor

Adaptor protein Shc undergoes translocation and mediates up‐regulation of the tyrosine kinase c‐Src in EGF‐stimulated A431 cells

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