Gab2 Phosphorylation by RSK Inhibits Shp2 Recruitment and Cell Motility

Zhang, Xiaocui; Lavoie, Geneviève; Fort, Loïc; Huttlin, Edward L.; Tcherkezian, Joseph; Galán, Jacob A.; Gu, Haihua; Gygi, Steven P.; Carréno, Sébastien; Roux, Philippe P.

doi:10.1128/mcb.01353-12

Cited by 30 publications

(27 citation statements)

References 44 publications

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“…This is in contrast to a previous study that indicated that ERK1/2-mediated Gab2 phosphorylation on Ser612 inhibits Shp2 recruitment (27). These discrepancies may be explained by results showing that the ERK1/2-activated protein kinase RSK directly phosphorylates Gab2 and thereby inhibits Shp2 recruitment (14). In this scenario, it may not be the direct action of ERK1/2 but rather RSK-dependent Gab2 phosphorylation that regulates Shp2 recruitment.…”

Section: Discussioncontrasting

confidence: 88%

“…Next, we sought to delineate the protein kinase(s) involved in Gab2 phosphorylation using pharmacological inhibitors of MEK1/2 (PD184352 and U0126) to prevent ERK1/2 activation. We also tested an RSK (p90 ribosomal S6 kinase) inhibitor (BI-D1870), which was previously shown to regulate Gab2 phosphorylation in response to agonists of the Ras/ERK pathway (14). While RSK inhibition modestly affected the mobility shift of Gab2 induced by PMA, we found that treatment of cells with MEK1/2 inhibitors efficiently blocked Gab2 phosphorylation (Fig.…”

Section: Resultsmentioning

confidence: 90%

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Extracellular Signal-Regulated Kinases 1 and 2 Phosphorylate Gab2 To Promote a Negative-Feedback Loop That Attenuates Phosphoinositide 3-Kinase/Akt Signaling

Zhang

Lavoie

Méant

et al. 2017

Molecular and Cellular Biology

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The scaffolding adapter protein Gab2 (Grb2-associated binder) promotes cell proliferation, survival, and motility by engaging several signaling pathways downstream of growth factor and cytokine receptors. In particular, Gab2 plays essential roles in mast cells, as it is required for phosphoinositide 3-kinase (PI3K) activation in response to Kit and the high-affinity IgE receptor. While the positive role of Gab2 in PI3K signaling is well documented, very little is known about the mechanisms that attenuate its function. Here we show that Gab2 becomes phosphorylated on multiple proline-directed sites upon stimulation of the Ras/extracellular signal-regulated kinase (ERK) signaling pathway. We demonstrate that ERK1 and ERK2 interact with Gab2 via a novel docking motif, which is required for subsequent Gab2 phosphorylation in response to ERK1/2 activation. We identified four ERK1/2-dependent phosphorylation sites in Gab2 that prevent the recruitment of the p85 regulatory subunit of PI3K. Using bone marrow-derived mast cells to study Gab2-dependent signaling, we found that the inhibition of ERK1/2 activity promotes Akt signaling in response to Kit and the high-affinity IgE receptor. Together, our results indicate that ERK1/2 participates in a negative-feedback loop that attenuates PI3K/Akt signaling in response to various agonists.

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Section: Discussioncontrasting

confidence: 88%

Section: Resultsmentioning

confidence: 90%

Extracellular Signal-Regulated Kinases 1 and 2 Phosphorylate Gab2 To Promote a Negative-Feedback Loop That Attenuates Phosphoinositide 3-Kinase/Akt Signaling

Zhang

Lavoie

Méant

et al. 2017

Molecular and Cellular Biology

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“…Taken together, even high doses of DST have minimal impact on the three PI3K dependent phosphorylation sites on Gab2 and its 14-3-3 binding potential. This is of particular interest as a recent study, primarily conducted in HEK293 cells, provided convincing evidence that the ERK/RSK axis can drive the phosphorylation of S159 and S210 as well [39]. However, the fact that we still observe substantial amounts of S159 and S210 phosphorylation despite the drastic loss of ERK phosphorylation suggests that the contribution of the ERK pathway plays a minor role in phosphorylation of these sites in K562 cells.…”

Section: Resultsmentioning

confidence: 52%

Alterations of Gab2 signalling complexes in imatinib and dasatinib treated chronic myeloid leukaemia cells

et al. 2013

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BackgroundThe Gab2 docking protein acts as an important signal amplifier downstream of various growth factor receptors and Bcr-Abl, the driver of chronic myeloid leukaemia (CML). Despite the success of Bcr-Abl tyrosine kinase inhibitors (TKI) in the therapy of CML, TKI-resistance remains an unsolved problem in the clinic. We have recently shown that Gab2 signalling counteracts the efficacy of four distinct Bcr-Abl inhibitors. In the course of that project, we noticed that two clinically relevant drugs, imatinib and dasatinib, provoke distinct alterations in the electrophoretic mobility of Gab2, its signalling output and protein interactions. As the signalling potential of the docking protein is highly modulated by its phosphorylation status, we set out to obtain more insights into the impact of TKIs on Gab2 phosphorylation.FindingsUsing stable isotope labelling by amino acids in cell culture (SILAC)-based quantitative mass spectrometry (MS), we show now that imatinib and dasatinib provoke distinct effects on the phosphorylation status and interactome of Gab2. This study identifies several new phosphorylation sites on Gab2 and confirms many sites previously known from other experimental systems. At equimolar concentrations, dasatinib is more effective in preventing Gab2 tyrosine and serine/threonine phosphorylation than imatinib. It also affects the phosphorylation status of more residues than imatinib. In addition, we also identify novel components of the Gab2 signalling complex, such as casein kinases, stathmins and PIP1 as well as known interaction partners whose association with Gab2 is disrupted by imatinib and/or dasatinib.ConclusionsBy using MS-based proteomics, we have identified new and confirmed known phosphorylation sites and interaction partners of Gab2, which may play an important role in the regulation of this docking protein. Given the growing importance of Gab2 in several tumour entities we expect that our results will help to understand the complex regulation of Gab2 and how this docking protein can contribute to malignancy.

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“…RSKs phosphorylate many proteins, both cytosolic and nuclear (2). The many effects of RSKs on various proteins may contribute to the observations that RSKs mediate wide-ranging cellular processes, including proliferation (6–8), migration (9), and invasion (1). …”

Section: Introductionmentioning

confidence: 99%

RSK Promotes Prostate Cancer Progression in Bone through ING3, CKAP2, and PTK6-Mediated Cell Survival

Lee

Cheng

et al. 2015

Molecular Cancer Research

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Prostate cancer (PCa) has a proclivity to metastasize to bone. The mechanism by which PCa cells are able to survive and progress in the bone microenvironment is not clear. Identification of molecules that play critical roles in the progression of PCa in bone will provide essential targets for therapy. Ribosomal S6 protein kinase (RSKs) have been shown to mediate many cellular functions critical for cancer progression. Whether RSK plays a role in the progression of PCa in bone is unknown. Immunohistochemical (IHC) analysis of human PCa specimens showed increased phosphorylation of RSK in the nucleus of PCa cells in a significant fraction of human PCa bone metastasis specimens, compared to the primary site or lymph node metastasis. Expression of constitutively active myristylated-RSK in C4-2B4 cells (C4-2B4/RSK) increased their survival and anchorage-independent growth compared to C4-2B4/vector cells. Using an orthotopic bone injection model, it was determined that injecting C4-2B4/RSK cells into mouse femurs enhanced their progression in bone compared to control cells. In PC3-mm2 cells, knockdown of RSK1 (RPS6KA1), the predominant RSK isoform, but not RSK2 (RPS6KA2) alone, decreased anchorage-independent growth in vitro and reduced tumor progression in bone and tumor-induced bone remodeling in vivo. Mechanistic studies showed that RSK regulates anchorage-independent growth through transcriptional regulation of factors that modulate cell survival, including ING3, CKAP2 and PTK6. Together, these data provide strong evidence that RSK is an important driver in PCa progression in bone. Implications RSK, an important driver in PCa progression in bone, has promising potential as a therapeutic target for PCa bone metastasis.

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Gab2 Phosphorylation by RSK Inhibits Shp2 Recruitment and Cell Motility

Cited by 30 publications

References 44 publications

Extracellular Signal-Regulated Kinases 1 and 2 Phosphorylate Gab2 To Promote a Negative-Feedback Loop That Attenuates Phosphoinositide 3-Kinase/Akt Signaling

Extracellular Signal-Regulated Kinases 1 and 2 Phosphorylate Gab2 To Promote a Negative-Feedback Loop That Attenuates Phosphoinositide 3-Kinase/Akt Signaling

Alterations of Gab2 signalling complexes in imatinib and dasatinib treated chronic myeloid leukaemia cells

RSK Promotes Prostate Cancer Progression in Bone through ING3, CKAP2, and PTK6-Mediated Cell Survival

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