The Phosphorylated 1169-Tyrosine Containing Region of Flt-1 Kinase (VEGFR-1) Is a Major Binding Site for PLCγ

Sawano, Asako; Takahashi, Tomoko; Yamaguchi, Sachiko; Shibuya, Masabumi

doi:10.1006/bbrc.1997.7327

Cited by 113 publications

(77 citation statements)

References 20 publications

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“…We have previously demonstrated the VEGF-dependent phosphorylation of PLC␥ in Flt1-expressing NIH3T3 cells (17). We also found that PLC␥ was phosphorylated on VEGF-treatment in AG1-G1-Flt1 cells.…”

Section: Discussionsupporting

confidence: 54%

“…Next, the potential role of RACK1 in this Flt1-dependent activation of the PI3K/Akt cascade was examined. As shown in Previous studies revealed that the phosphorylation of 1169-Tyr of Flt1 corresponding to 1175-Tyr of KDR relays a signal to PLC␥ and MAPK, leading to the proliferation of endothelial cells (15)(16)(17)32). Hence, the effect of RACK1-silencing on the VEGF-triggered activation of PLC␥ and MAPK was examined.…”

Section: Rack1-silencing Suppresses the Vegf-driven Activation Of Thementioning

confidence: 93%

“…The phosphorylation of Tyr(Y)-1175 on KDR leads to the activation of phospholipase C (PLC)␥, which in turn promotes the intracellular mobilization of calcium and activates a crucial protein kinase C-Raf-mitogen-activated protein kinase (PKC-Raf-MAPK) cascade, the latter regulating endothelial cell proliferation (14 -16). The phosphorylation of Tyr(Y)-1169 on Flt1 also provides a binding site for PLC␥ and activates a PLC␥-MAPK cascade (17). Moreover, both receptors appear to activate the PI3 kinase (PI3K)-Akt pathway (18,19).…”

mentioning

confidence: 99%

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RACK1 Regulates VEGF/Flt1-mediated Cell Migration via Activation of a PI3K/Akt Pathway

Wang

Yamauchi

Matsushima

et al. 2011

Journal of Biological Chemistry

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Vascular endothelial growth factor (VEGF) is vital to physiological as well as pathological angiogenesis, and regulates a variety of cellular functions, largely by activating its 2 receptors, fms-like tyrosine kinase (Flt1) and kinase domain receptor (KDR). KDR plays a critical role in the proliferation of endothelial cells by controlling VEGF-induced phospholipase C␥-protein kinase C (PLC␥-PKC) signaling. The function of Flt1, however, remains to be clarified. Recent evidence has indicated that Flt1 regulates the VEGF-triggered migration of endothelial cells and macrophages. Here, we show that RACK1, a ubiquitously expressed scaffolding protein, functions as an important regulator of this process. We found that RACK1 (receptor for activated protein kinase C 1) binds to Flt1 in vitro. When the endogenous expression of RACK1 was attenuated by RNA interference, the VEGF-driven migration was remarkably suppressed whereas the proliferation was unaffected in a stable Flt1-expressing cell line, AG1-G1-Flt1. Further, we demonstrated that the VEGF/Flt-mediated migration of AG1-G1-Flt1 cells occurred mainly via the activation of the PI3 kinase (PI3K)/ Akt and Rac1 pathways, and that RACK1 plays a crucial regulatory role in promoting PI3K/Akt-Rac1 activation.

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Section: Discussionsupporting

confidence: 54%

Section: Rack1-silencing Suppresses the Vegf-driven Activation Of Thementioning

confidence: 93%

mentioning

confidence: 99%

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RACK1 Regulates VEGF/Flt1-mediated Cell Migration via Activation of a PI3K/Akt Pathway

Wang

Yamauchi

Matsushima

et al. 2011

Journal of Biological Chemistry

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“…Among VEGF receptors, KDR/Flk-1 appears to be responsible for most of this signaling, since Flt-1 carries a very weak tyrosine kinase activity (Seetharam et al, 1995;Waltenberger et al, 1994;Sawano et al, 1997).…”

Section: Discussionmentioning

confidence: 99%

VEGF activates protein kinase C-dependent, but Ras-independent Raf-MEK-MAP kinase pathway for DNA synthesis in primary endothelial cells

1999

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“…In several growth factor receptors a specific autophosphorylation site is essential for PLC-␥1 association: Tyr-1021 in the platelet-derived growth factor ␤-receptor (19, 24 -26), Tyr-766 in the fibroblast growth factor receptor (17,18), Tyr-785 in the nerve growth factor receptor Trk (27,28), Tyr-1169 in the vascular endothelial growth factor receptor Flt (29), Tyr-1015 in the glial-derived growth factor receptor Ret (30,31), and Tyr-1356 of the hepatocyte growth factor receptor Met (32,33). In each instance, mutagenesis of the essential tyrosine produces near complete abrogation of PLC-␥1 association and tyrosine phosphorylation.…”

mentioning

confidence: 99%

The Role of Individual SH2 Domains in Mediating Association of Phospholipase C-γ1 with the Activated EGF Receptor

Chattopadhyay¹,

Vecchi²,

Ji³

et al. 1999

Journal of Biological Chemistry

131

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The two SH2 (Src homology domain 2) domains present in phospholipase C-␥1 (PLC-␥1) were assayed for their capacities to recognize the five autophosphorylation sites in the epidermal growth factor receptor. Plasmon resonance and immunological techniques were employed to measure interactions between SH2 fusion proteins and phosphotyrosine-containing peptides. The N-SH2 domain recognized peptides in the order of pY1173 > pY992 > pY1068 > pY1148 > > pY1086, while the C-SH2 domain recognized peptides in the order of pY992 > pY1068 > pY1148 > > pY1086 and pY1173. The major autophosphorylation site, pY1173, was recognized only by the N-SH2 domain. Contributions of the N-SH2 and C-SH2 domains to the association of the intact PLC-␥1 molecule with the activated epidermal growth factor (EGF) receptor were assessed in vivo. Loss of function mutants of each SH2 domain were produced in a full-length epitope-tagged PLC-␥1. After expression of the mutants, cells were treated with EGF and association of exogenous PLC-␥1 with EGF receptors was measured. In this context the N-SH2 is the primary contributor to PLC-␥1 association with the EGF receptor. The combined results suggest an association mechanism involving the N-SH2 domain and the pY1173 autophosphorylation site as a primary event and the C-SH2 domain and the pY992 autophosphorylation site as a secondary event.A rapid cellular response to growth factor binding to cell surface receptors is the hydrolysis of phosphatidylinositol 4,5-bisphosphate to produce two second messengers: inositol 1,4,5-trisphosphate and diacylglycerol (1). Respectively, these molecules initiate the mobilization of intracellular Ca 2ϩ and activation of protein kinase C. The mechanism by which growth factors stimulate this reaction involves the tyrosine phosphorylation-dependent activation of a specific phospholipase C (PLC) isoform: PLC-␥1 or PLC-␥2 (2, 3). Other PLC isoforms, PLC 1 -␤1-4 or PLC-␦1-4, are activated by growth factor-independent or unknown mechanisms. PLC-␥1 is ubiquitously expressed and targeted disruption of its gene in mice results in embryonic lethality (4). The PLC-␥2 species has a more restricted distribution (5) and has been investigated less. A PLC-␥ gene has been identified in Drosophila and its disruption leads to aberrant eye development (6). In cell culture systems, however, the role of PLC-␥1 in the mitogenic response is unclear and has been described as both essential (7-13) and non-essential (14 -20).PLC-␥ isoforms are structurally unique within this phospholipase family as they include Src homology (SH) domains, which enable protein/protein interactions (21, 22). PLC-␥1 contains two SH2 domains, which are 35% identical in amino acid sequence, and one SH3 domain. The SH2 domains mediate the association of PLC-␥1 with autophosphorylation sites on activated receptor tyrosine kinases (23), an essential prerequisite to PLC-␥1 tyrosine phosphorylation and activation. The function of SH3 domains is to mediate association with proline-rich sequences in partner proteins; however, ...

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The Phosphorylated 1169-Tyrosine Containing Region of Flt-1 Kinase (VEGFR-1) Is a Major Binding Site for PLCγ

Cited by 113 publications

References 20 publications

RACK1 Regulates VEGF/Flt1-mediated Cell Migration via Activation of a PI3K/Akt Pathway

RACK1 Regulates VEGF/Flt1-mediated Cell Migration via Activation of a PI3K/Akt Pathway

VEGF activates protein kinase C-dependent, but Ras-independent Raf-MEK-MAP kinase pathway for DNA synthesis in primary endothelial cells

The Role of Individual SH2 Domains in Mediating Association of Phospholipase C-γ1 with the Activated EGF Receptor

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