Two distinct pools of Src family tyrosine kinases regulate PDGF-induced DNA synthesis and actin dorsal ruffles

Veracini, Laurence; Franco, Mélanie; Boureux, Anthony; Simon, Valérie; Roche, Serge; Bénistant, Christine

doi:10.1242/jcs.03015

Cited by 47 publications

(65 citation statements)

References 64 publications

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“…These data show that all agents activated Src. Although the magnitude of the increase in phosphorylation was modest, it is consistent with what other groups report using this approach (30,64) and what we have found using a different approach to monitor activation of SFKs (43). B, mouse embryo fibroblasts that lacked all SFKs (39) were reconstituted with either an empty expression vector (Ϫ/Ϫ), or wild type c-Src (Src) (40) and stimulated with the indicated agents as described in panel A.…”

Section: Indirect Activation Of Pdgfr␣ Promoted Proliferation and Sursupporting

confidence: 91%

Growth Factors Outside of the Platelet-derived Growth Factor (PDGF) Family Employ Reactive Oxygen Species/Src Family Kinases to Activate PDGF Receptor α and Thereby Promote Proliferation and Survival of Cells

Lei

Kazlauskas

2009

Journal of Biological Chemistry

100

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The vitreous contains a plethora of growth factors that are strongly implicated in the formation of fibroproliferative diseases such as proliferative vitreoretinopathy. Although plateletderived growth factors (PDGFs) are present in the vitreous, vitreal growth factors outside of the PDGF family activated the PDGF ␣ receptor (PDGFR␣) and promoted disease progression in a rabbit model of proliferative vitreoretinopathy (H. Lei, G. Velez, P. Hovland, T. Hirose, D. Gilbertson, and A. Kazlauskas (2008) submitted for publication.) In this report we investigated the mechanism by which non-PDGFs activated PDGFR␣. We found that non-PDGFs increased the cellular level of reactive oxygen species (ROS) and that this event was necessary and sufficient for phosphorylation of PDGFR␣. We speculated that the underlying mechanism was ROS-mediated inhibition of phosphotyrosine phosphatases, which antagonize receptor autophosphorylation. However, this did not appear to be the case. Non-PDGFs promoted tyrosine phosphorylation of catalytically inactive PDGFR␣, and thereby indicated that at least one additional tyrosine kinase was involved. Indeed, preventing expression or blocking the kinase activity of Src family kinases suppressed non-PDGF-dependent tyrosine phosphorylation of PDGFR␣. Thus non-PDGFs increased the level of ROS, which activated Src family kinases and resulted in phosphorylation of PDGFR␣. Finally, although non-PDGFs induced only modest phosphorylation of PDGFR␣, proliferation and survival of cells in response to non-PDGFs was significantly enhanced by expression of PDGFR␣. These studies reveal a novel mechanism for activation of PDGFR␣ that appears capable of enhancing the responsiveness of cells to growth factors outside of the PDGF family.

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Section: Indirect Activation Of Pdgfr␣ Promoted Proliferation and Sursupporting

confidence: 91%

Growth Factors Outside of the Platelet-derived Growth Factor (PDGF) Family Employ Reactive Oxygen Species/Src Family Kinases to Activate PDGF Receptor α and Thereby Promote Proliferation and Survival of Cells

Lei

Kazlauskas

2009

Journal of Biological Chemistry

100

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“…Ras and Rab5 pathways have also been documented (Lanzetti et al, 2004), and we have recently shown that this response implicates Src family tyrosine kinases (SFK). In addition, our results suggest that in contrast to Src mitogenic function, SFK signal transduction promoting actin assembly does not require receptor association (Veracini et al, 2006). Rather, it involves the lipid second messenger sphingosine-1-phosphate, which may activate SFK through a seven-transmembrane receptor of the EDG family and an associated heterotrimeric Gi protein.…”

mentioning

confidence: 74%

“…This potentiating effect was specific to dorsal ruffles as lateral ruffles were not (Manes et al, 2000) were subcloned into pBABE. Cell culture, transfection, infection, immunofluorescence analysis, Q-PCR and biochemistry have been described by Veracini et al (2006). White arrows indicate cells with dorsal ruffles.…”

Section: Slap Negatively Regulates Pdgf-induced Dorsal Rufflesmentioning

confidence: 99%

The Src-like adaptor protein regulates PDGF-induced actin dorsal ruffles in a c-Cbl-dependent manner

et al. 2008

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The Src-like adaptor protein (SLAP) belongs to the subfamily of adapter proteins that negatively regulate cellular signalling initiated by tyrosine kinases. SLAP has a unique, myristylated N-terminus, followed by SH3 and SH2 domains with high homology to Src family tyrosine kinases (SFK) and a unique C-terminal tail, which is important for c-Cbl binding. We have previously shown that SLAP negatively regulates platelet-derived growth factor (PDGF)-induced mitogenesis in fibroblasts and we now report that it regulates F-actin assembly for dorsal ruffles formation. c-Cbl mediated SLAP inhibition towards actin remodelling. Moreover, SLAP enhanced PDGF-induced c-Cbl phosphorylation by SFK. In contrast, SLAP mitogenic inhibition was not mediated by c-Cbl, but it rather involved a competitive mechanism with SFK for PDGF-receptor (PDGFR) association and mitogenic signalling. Accordingly, phosphorylation of the Src mitogenic substrates Stat3 and Shc were reduced by SLAP. Thus, we concluded that SLAP regulates PDGFR signalling by two independent mechanisms: a competitive mechanism for PDGF-induced Src mitogenic signalling and a non-competitive mechanism for dorsal ruffles formation mediated by c-Cbl.

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“…Moreover, Csk relocalization to these membranes induced, at most, a 50% reduction of SFK activity, confirming that it does not target all pools of SFK present in these cells. Accordingly, spatial regulation has been considered as an important feature of signal specificity induced by SFK in nontransformed cells (Veracini et al, 2006(Veracini et al, , 2008, and this may apply for transformed cells as well. For example, when localized at focal adhesions, SFK has important roles in cell growth and survival (Frame, 2004).…”

Section: Discussionmentioning

confidence: 99%

“…Purified FynK298M (FynK À ) was described in Koegl et al (1994). Anti-SFK (cst1) was described in Veracini et al (2006); anti-Src specific (2.17) antibody was a generous gift from Dr S Parsons (University of Virginia, VA, USA). Anti-Csk was described in Be´nistant et al (2001), anti-PAG in Veracini et al (2008) and anti-Cbp in Oneyama et al (2008).…”

Section: Methodsmentioning

confidence: 99%

Src family tyrosine kinases-driven colon cancer cell invasion is induced by Csk membrane delocalization

et al. 2009

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The nonreceptor tyrosine kinases of the Src family (SFK) are frequently deregulated in human colorectal cancer (CRC), and they have been implicated in tumour growth and metastasis. How SFK are activated in this cancer has not been clearly established. Here, we show that the SFK-dependent invasion is induced by inactivation of the negative regulator C-terminal Src kinase, Csk. While the level of Csk was inconsistent with SFK activity in colon cancer cells, its membrane translocation, needed for efficient regulation of membrane-localized SFK activity, was impaired. Accordingly, Csk downregulation did not affect SFK oncogenic activity in these cells, whereas expression of a membrane-localized form of this kinase affected their invasive activity. Downregulation of the transmembrane and rafts-localized Csk-binding protein/ phosphoprotein associated with glycosphingolipid-enriched microdomain (PAG), was instrumental for the cytoplasmic accumulation of Csk. Re-expression of PAG in cells from late-stage CRC inhibited SFK invasive activity in a Cskdependent manner. Conversely, inactivation of its residual expression in early-stage CRC cells promoted SFK invasive activity. Finally, this mechanism was specific to CRC as Csk coupling to SFK was readily detected in breast cancer cells. Therefore, Csk mis-localization defines a novel mechanism for SFK oncogenic activation in CRC cells.

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Two distinct pools of Src family tyrosine kinases regulate PDGF-induced DNA synthesis and actin dorsal ruffles

Cited by 47 publications

References 64 publications

Growth Factors Outside of the Platelet-derived Growth Factor (PDGF) Family Employ Reactive Oxygen Species/Src Family Kinases to Activate PDGF Receptor α and Thereby Promote Proliferation and Survival of Cells

Growth Factors Outside of the Platelet-derived Growth Factor (PDGF) Family Employ Reactive Oxygen Species/Src Family Kinases to Activate PDGF Receptor α and Thereby Promote Proliferation and Survival of Cells

The Src-like adaptor protein regulates PDGF-induced actin dorsal ruffles in a c-Cbl-dependent manner

Src family tyrosine kinases-driven colon cancer cell invasion is induced by Csk membrane delocalization

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