The control of phosphatidylinositol 3,4-bisphosphate concentrations by activation of the Src homology 2 domain containing inositol polyphosphate 5-phosphatase 2, SHIP2

Activation of class Ia PI3K (phosphoinositide 3-kinase) produces PtdInsP3, a vital intracellular mediator whose degradation generates additional lipid signals. In the present study vanadate analogues that inhibit PTPs (protein tyrosine phosphatases) were used to probe the mechanisms which regulate the concentrations of these molecules allowing their independent or integrated function. In 1321N1 cells, which lack PtdInsP3 3-phosphatase activity, sodium vanadate or a cell permeable derivative, bpV(phen) [potassium bisperoxo(1,10-phenanthroline)oxovanadate (V)], increased the recruitment into anti-phosphotyrosine immunoprecipitates of PI3K activity and of the p85 and p110α subunits of class Ia PI3K and enhanced the recruitment of PI3K activity stimulated by PDGF (platelet-derived growth factor). However, neither inhibitor much increased cellular PtdInsP3 concentrations, but both diminished dramatically the accumulation of PtdInsP3 stimulated by PDGF or insulin and markedly increased the control and stimulated concentrations of PtdIns(3,4)P2. These actions were accounted for by the ability of PTP inhibitors to stimulate the activity of endogenous PtdInsP3 5-phosphatase(s), particularly SHIP2 (Src homology 2 domain containing inositol polyphosphate 5-phosphatase 2) and to inhibit types I and II PtdIns(3,4)P2 4-phosphatases. Thus bpV(phen) promoted the translocation of SHIP2 from the cytosol to a Triton X-100-insoluble fraction and induced a marked (5–10-fold) increase in SHIP2 specific activity mediated by enhanced tyrosine phosphorylation. The net effect of these inhibitors was, therefore, to switch the signal output of class I PI3K from PtdInsP3 to PtdIns(3,4)P2. A key component controlling this shift in the balance of lipid signals is the activation of SHIP2 by increased tyrosine phosphorylation, an effect observed in HeLa cells in response to both PTP inhibitors and epidermal growth factor.

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Direct analysis of the binding of the abl Src homology 2 domain to the activated epidermal growth factor receptor.

Journal of Biological Chemistry ◽

10.1016/s0021-9258(18)53920-x ◽

1993 ◽

Vol 268 (3) ◽

pp. 1775-1779 ◽

Cited By ~ 1

Author(s):

G. Zhu ◽

S.J. Decker ◽

B.J. Mayer ◽

A.R. Saltiel

Keyword(s):

Epidermal Growth Factor Receptor ◽

Growth Factor ◽

Epidermal Growth Factor ◽

Growth Factor Receptor ◽

Direct Analysis ◽

Src Homology 2 ◽

Src Homology ◽

Epidermal Growth ◽

Src Homology 2 Domain

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Nuclear Magnetic Resonance Solution Structure of the Growth Factor Receptor-Bound Protein 2 Src Homology 2 Domain

Biochemistry ◽

10.1021/bi952615s ◽

1996 ◽

Vol 35 (36) ◽

pp. 11852-11864 ◽

Cited By ~ 12

Author(s):

Kevin H. Thornton ◽

W. Tom Mueller ◽

Patrick McConnell ◽

Guochang Zhu ◽

Alan R. Saltiel ◽

...

Keyword(s):

Nuclear Magnetic Resonance ◽

Growth Factor ◽

Magnetic Resonance ◽

Solution Structure ◽

Growth Factor Receptor ◽

Src Homology 2 ◽

Resonance Solution ◽

Src Homology ◽

Src Homology 2 Domain ◽

Growth Factor Receptor Bound

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Src Homology 2 Domain-Containing Inositol Polyphosphate Phosphatase Regulates NF-κB-Mediated Gene Transcription by Phagocytic FcγRs in Human Myeloid Cells

The Journal of Immunology ◽

10.4049/jimmunol.169.8.4370 ◽

2002 ◽

Vol 169 (8) ◽

pp. 4370-4378 ◽

Cited By ~ 44

Author(s):

Susheela Tridandapani ◽

Yijie Wang ◽

Clay B. Marsh ◽

Clark L. Anderson

Keyword(s):

Gene Transcription ◽

Myeloid Cells ◽

Inositol Polyphosphate ◽

Src Homology 2 ◽

Src Homology ◽

Src Homology 2 Domain

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SRC-homology 2 domain-containing phosphatase 2 (SHP2) in epidermal growth factor receptor (EGFR)-mutant lung adenocarcinoma (LUAD).

Journal of Clinical Oncology ◽

10.1200/jco.2020.38.15_suppl.9540 ◽

2020 ◽

Vol 38 (15_suppl) ◽

pp. 9540-9540

Author(s):

Rafael Rosell ◽

Masaoki Ito ◽

Jordi Codony-Servat ◽

Ana Giménez-Capitán ◽

Mireia Serra-Mitjans ◽

...

Keyword(s):

Growth Factor ◽

Epidermal Growth Factor ◽

Mrna Expression ◽

Mrna Levels ◽

Wild Type ◽

Src Homology 2 ◽

Src Homology ◽

Epidermal Growth ◽

Src Homology 2 Domain ◽

Egfr Mutant

9540 Background: Epidermal growth factor (EGFR)-mutant lung adenocarcinomas (LUADs) display impaired phosphorylation of extracellular signal-regulated kinase (ERK) and SRC-homology 2 domain-containing phosphatase 2 (SHP2) in comparison with EGFR wild-type LUADs. However, the function of SHP2 in early EGFR-mutant LUADs and EGFR wild-type LUADs has not been reported. We posit that SHP2 mRNA expression could be a predictive marker in resected EGFR-mutant LUADs versus EGFR wild-type patients (pts). Methods: We examined 267 resected LUADs from Japan and Spain. mRNA expression levels of AXL, MET, CDCP1, STAT3, YAP1 and SHP2 were analyzed by quantitative reverse transcriptase polymerase chain reaction (PCR). EGFR mutant cell lines were investigated for their activity of SHP2. Results: Among the 267 enrolled pts, 100 (37.3%) were EGFR-mutant LUADs. Five-year recurrence-free survival (RFS) and overall survival (OS) were lower for EGFR-mutant LUADs with high SHP2 mRNA levels (hazard ratio = 1.83 and 2.28, respectively. p = 0.03 and p = 0.04). However, SHP2 was not associated with RFS nor OS in the 167 wild-type EGFR LUADs. In EGFR-mutant cells, RMC-4550 (SHP2 inhibitor) plus erlotinib showed synergism via inhibition of AKT (S473) and ERK1/2 (T202/Y204). While erlotinib translocates SHP2 (Y542) into the nucleus, either RMC-4550 alone, or in combination with erlotinib, relocalizes SHP2 into the cytoplasm membrane, limiting AKT and ERK activation. Conclusions: High SHP2 mRNA is related to shorter RFS and OS in EGFR-mutant LUADs, but not in EGFR wild-type LUADs. The findings indicate that the addition of SHP2 inhibitors could improve adjuvant therapy in EGFR-mutant LUADs.

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Selectivity and Mechanism of Action of a Growth Factor Receptor-Bound Protein 2 Src Homology 2 Domain Binding Antagonist

Journal of Medicinal Chemistry ◽

10.1021/jm800523u ◽

2008 ◽

Vol 51 (23) ◽

pp. 7459-7468 ◽

Cited By ~ 7

Author(s):

Alessio Giubellino ◽

Zhen-Dan Shi ◽

Lisa M. Miller Jenkins ◽

Karen M. Worthy ◽

Lakshman K. Bindu ◽

...

Keyword(s):

Growth Factor ◽

Mechanism Of Action ◽

Growth Factor Receptor ◽

Src Homology 2 ◽

Src Homology ◽

Src Homology 2 Domain ◽

Growth Factor Receptor Bound

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Multiple stimuli induce tyrosine phosphorylation of the Crk-binding sites of paxillin

Biochemical Journal ◽

10.1042/bj3600057 ◽

2001 ◽

Vol 360 (1) ◽

pp. 57-66 ◽

Cited By ~ 19

Author(s):

Michael D. SCHALLER ◽

Erik M. SCHAEFER

Keyword(s):

Cell Adhesion ◽

Tyrosine Phosphorylation ◽

Binding Sites ◽

Signalling Molecules ◽

Src Homology 2 ◽

Dependent Cell ◽

Src Homology ◽

Tyrosine Phosphorylated Protein ◽

Src Homology 2 Domain

Paxillin is a focal-adhesion-associated, tyrosine-phosphorylated protein. In cells transformed by the src, crk or BCR-Abl oncogenes, the phosphotyrosine content of paxillin is elevated. In normal cells paxillin functions in signalling following integrin-dependent cell adhesion or exposure to a number of stimuli, including growth factors and neuropeptides. These stimuli induce tyrosine phosphorylation of paxillin, regulating the association of Src homology 2 domain-containing signalling molecules with paxillin. There are multiple sites of tyrosine phosphorylation on paxillin. To elucidate the role of paxillin in transducing signals in response to various stimuli, it is essential to identify all of the sites of phosphorylation on paxillin and to define which residues are phosphorylated in response to distinct stimuli. We describe two new sites of tyrosine phosphorylation on paxillin, residues at positions 40 and 88. Using paxillin variants with phenylalanine substitutions at phosphorylation sites and phospho-specific paxillin antibodies, tyrosine phosphorylation of paxillin in response to distinct stimuli was examined. The results demonstrate that Tyr31 and Tyr118, which are binding sites for Crk, are major sites of tyrosine phosphorylation following cell adhesion or stimulation with platelet-derived growth factor or angiotensin II. Thus multiple stimuli may elicit similar signalling events downstream of paxillin.

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Interdependent fibroblast growth factor and activin A signaling promotes the expression of endodermal genes in differentiating mouse embryonic stem cells expressing Src Homology 2-domain inactive Shb

Differentiation ◽

10.1111/j.1432-0436.2007.00249.x ◽

2008 ◽

Vol 76 (5) ◽

pp. 443-453 ◽

Cited By ~ 8

Author(s):

Nina S. Funa ◽

Johan Saldeen ◽

Björn Åkerblom ◽

Michael Welsh

Keyword(s):

Stem Cells ◽

Embryonic Stem Cells ◽

Growth Factor ◽

Fibroblast Growth Factor ◽

Embryonic Stem ◽

Activin A ◽

Fibroblast Growth ◽

Src Homology 2 ◽

Src Homology ◽

Src Homology 2 Domain

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B Cell Adaptor Containing Src Homology 2 Domain (Bash) Links B Cell Receptor Signaling to the Activation of Hematopoietic Progenitor Kinase 1

Journal of Experimental Medicine ◽

10.1084/jem.194.4.529 ◽

2001 ◽

Vol 194 (4) ◽

pp. 529-540 ◽

Cited By ~ 48

Author(s):

Sachiyo Tsuji ◽

Mariko Okamoto ◽

Koichi Yamada ◽

Noriaki Okamoto ◽

Ryo Goitsuka ◽

...

Keyword(s):

B Cells ◽

B Cell ◽

Tyrosine Phosphorylation ◽

Phosphorylation Site ◽

Sh2 Domain ◽

Hematopoietic Progenitor ◽

Src Homology 2 ◽

Bcr Signaling ◽

Src Homology ◽

Src Homology 2 Domain

The B cell adaptor containing src homology 2 domain (BASH; also termed BLNK or SLP-65), is crucial for B cell antigen receptor (BCR)-mediated activation, proliferation, and differentiation of B cells. BCR-mediated tyrosine-phosphorylation of BASH creates binding sites for signaling effectors such as phospholipase Cγ (PLCγ)2 and Vav, while the function of its COOH-terminal src homology 2 domain is unknown. We have now identified hematopoietic progenitor kinase (HPK)1, a STE20-related serine/threonine kinase, as a protein that inducibly interacts with the BASH SH2 domain. BCR ligation induced rapid tyrosine-phosphorylation of HPK1 mainly by Syk and Lyn, resulting in its association with BASH and catalytic activation. BCR-mediated activation of HPK1 was impaired in Syk- or BASH-deficient B cells. The functional SH2 domain of BASH and Tyr-379 within HPK1 which we identified as a Syk-phosphorylation site were both necessary for interaction of both proteins and efficient HPK1 activation after BCR stimulation. Furthermore, HPK1 augmented, whereas its kinase-dead mutant inhibited IκB kinase β (IKKβ) activation by BCR engagement. These results reveal a novel BCR signaling pathway leading to the activation of HPK1 and subsequently IKKβ, in which BASH recruits tyrosine-phosphorylated HPK1 into the BCR signaling complex.

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