Differential involvement of Src family kinases in pervanadate-mediated responses in rat myometrial cells

Boulven, Isaline; Robin, Philippe; Desmyter, Christine; Harbon, Simone; Leiber, Denis

doi:10.1016/s0898-6568(01)00269-8

Cited by 20 publications

(27 citation statements)

References 31 publications

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“…PI-PLC phosphorylation is required for activation. [23][24][25] Our results show that though DEX treatment does not influence Src/PI-PLC complex formation, it does induce PI-PLC phosphorylation and activation. PI-PLC phosphorylation and apoptosis are inhibited by PP1 and by the specific PI-PLC inhibitor U73122, but not by the PC-PLC inhibitor D609.…”

Section: Discussionmentioning

confidence: 60%

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Dexamethasone-induced apoptosis of thymocytes: role of glucocorticoid receptor–associated Src kinase and caspase-8 activation

Marchetti

Marco

Cifone

et al. 2003

Blood

115

116

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Glucocorticoid hormones (GCHs) regulate normal and neoplastic lymphocyte development by exerting antiproliferative and/or apoptotic effects. We have previously shown that dexamethasone (DEX)-activated thymocyte apoptosis requires a sequence of events including interaction with the glucocorticoid receptor (GR), phosphatidylinositol-specific phospholipase C (PI-PLC), and acidic sphingomyelinase (aSMase) activation. We analyzed the mechanisms of GCH-activated apoptosis by focusing on GR-associated Src kinase, cytochrome c release, and caspase-8, -9, and -3 activation. We show here that PI-PLC binds to GR-associated Src kinase, as indicated by coimmunoprecipitation experiments. Moreover, DEX treatment induces PI-PLC phosphorylation and activation. DEX-induced PI-PLC phosphorylation, activation, and apoptosis are inhibited by PP1, a Src kinase inhibitor, thus suggesting that Src-mediated PI-PLC activation is involved in DEX-induced apoptosis. Caspase-9, -8, and -3 activation and cytochrome c release can be detected 1 to 2 hours after DEX treatment. Caspase-9 inhibition does not counter cytochrome c release, caspase-8 and caspase-3 activation, and apoptosis. Caspase-8 inhibition counters cytochrome c release, caspase-9 and caspase-3 activation, and apoptosis, thus suggesting that caspase-8 inhibitor can directly inhibit caspase-9 and/or that DEX-

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Section: Discussionmentioning

confidence: 60%

“…23 PI-PLC activation is reported to be a consequence of phosphorylation, which is blocked by PP1. 24,25 We evaluated whether DEX induces PI-PLC phosphorylation. In the same Src/PI-PLC coimmunoprecipitation experiment, we analyzed the presence of phosphorylated proteins.…”

Section: Pi-plc Associates With the Hsp90/gr/src Macromolecular Complmentioning

confidence: 99%

Dexamethasone-induced apoptosis of thymocytes: role of glucocorticoid receptor–associated Src kinase and caspase-8 activation

Marchetti

Marco

Cifone

et al. 2003

Blood

115

116

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“…Pervanadate enhances tyrosine phosphorylation by blocking dephosphorylation, and by activating tyrosine kinases (e.g. insulin receptor, EGFR, Src family kinases) regulated by tyrosine phosphorylation (Boulven et al, 2002;Fantus et al, 1989;Kim et al, 2003). Thus, PV-induced tyrosine phosphorylation probably reflects both ongoing physiological regulation and additional sites targeted by PV-activated kinases.…”

Section: Resultsmentioning

confidence: 99%

Tyrosine phosphorylation of nuclear-membrane protein emerin by Src, Abl and other kinases

Tifft

Bradbury

Wilson

2009

Journal of Cell Science

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X-linked recessive Emery-Dreifuss muscular dystrophy (EDMD) is caused by loss of emerin, a nuclear-membrane protein with roles in nuclear architecture, gene regulation and signaling. Phosphoproteomic studies have identified 13 sites of tyrosine phosphorylation in emerin. We validated one study, confirming that emerin is hyper-tyrosine-phosphorylated in Her2-overexpressing cells. We discovered that non-receptor tyrosine kinases Src and Abl each phosphorylate emerin and a related protein, LAP2, directly. Src phosphorylated emerin specifically at Y59, Y74 and Y95; the corresponding triple Yto-F ('FFF') mutation reduced tyrosine phosphorylation bỹ 70% in vitro and in vivo. Substitutions that removed a single hydroxyl moiety either decreased (Y19F, Y34, Y161F) or increased (Y4F) emerin binding to BAF in cells. Y19F, Y34F, Y161F and the FFF mutant also reduced recombinant emerin binding to BAF from HeLa lysates, demonstrating the involvement of both LEM-domain and distal phosphorylatable tyrosines in binding BAF. We conclude that emerin function is regulated by multiple tyrosine kinases, including Her2, Src and Abl, two of which (Her2, Src) regulate striated muscle. These findings suggest roles for emerin as a downstream effector and 'signal integrator' for tyrosine kinase signaling pathway(s) at the nuclear envelope.Key words: Emerin, LEM domain, Nuclear envelope, Emery-Dreifuss muscular dystrophy, Src, Her2, Abl, Barrier-to-autointegration factor (BAF), Cardiomyopathy, Breast cancer, Laminopathy, Neuromuscular junction Journal of Cell Science 3781 Emerin tyrosine phosphorylation receptor tyrosine kinases that can be activated by Her2 or other signaling pathways (Roskoski, 2005;Srinivasan and Plattner, 2006). We confirm Her2-stimulated emerin tyrosine phosphorylation and report that emerin and LAP2 are phosphorylated directly by Src and Abl. We also show that tyrosine phosphorylation of emerin regulates binding to BAF. ResultsWe compiled data from 13 independent proteomic studies (Amanchy et al., 2005;Brill et al., 2004;Cantin et al., 2008; Daub et al., 2008; Guo et al., 2008;Olsen et al., 2006;Pan et al., 2008;Rikova et al., 2007;Rush et al., 2005;Schlosser et al., 2006;Sui et al., 2008;Tao et al., 2005;Tsai et al., 2008), which identified 13 phosphorylated tyrosines in emerin (Fig. 1A). All sites except Y19 and Y41 were identified more than once. All 13 identified phosphorylated Tyr (Tyr-P) residues in human emerin are conserved in mouse, and six (corresponding to Y4, Y41, Y94, Y95, Y105, Y161) are conserved in Xenopus (Fig. 1B). These studies did not reveal responsible pathways or kinases.We validated tyrosine phosphorylation of emerin by immunoprecipitating endogenous emerin from HeLa cells treated with 1 M pervanadate (PV, a tyrosine phosphatase inhibitor) (Huyer et al., 1997) or buffer (PBS) as control, for 30 minutes before lysing cells. Emerin immunoprecipitated from PV-treated cells was specifically recognized by Tyr-P antibodies (not shown; see Fig. 2B). No other tyrosine-phosphorylated bands were dete...

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“…It is indeed possible that pervanadate may activate c-Src, which could phosphorylate connexin45 directly. To this point, c-Src and Src family members have been reported to be activated by pervanadate in a diverse variety of cells (Boulven et al, 2002;Fan et al, 2003;Takahashi et al, 2004;Heiska and Carpen, 2005). On the other hand, pervanadate inhibition of phosphatases may merely permit the detection of low level, constitutive tyrosine phosphorylation caused by currently unknown tyrosine kinases.…”

Section: D) Regulation Of Connexin43 Channel Gating By the Src Tyrosimentioning

confidence: 99%

Gap junction channel gating modulated through protein phosphorylation

Moreno

Lau

2007

Progress in Biophysics and Molecular Biology

106

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As a ubiquitous post-translation modification process, protein phosphorylation has proven to be a key mechanism in regulating the function of several membrane proteins, including transporters and channels. Connexins, pannexins, and innexins are protein families that form gap junction channels essential for intercellular communication. Connexins have been intensely studied, and most of their isoforms are known to be phosphorylated by protein kinases that lead to modifications in tyrosine, serine, and threonine residues, which have been reported to affect, in one way or another, intercellular communication. Despite the abundant reports on changes in intercellular communication due to the activation or inactivation of numerous kinases, the molecular mechanisms by which phosphorylation alters channel gating properties have not been elucidated completely. Hence, this chapter will cover some of the current, relevant research that attempt to explain how phosphorylation triggers and/or modulates gap junction channel gating.

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Differential involvement of Src family kinases in pervanadate-mediated responses in rat myometrial cells

Cited by 20 publications

References 31 publications

Dexamethasone-induced apoptosis of thymocytes: role of glucocorticoid receptor–associated Src kinase and caspase-8 activation

Dexamethasone-induced apoptosis of thymocytes: role of glucocorticoid receptor–associated Src kinase and caspase-8 activation

Tyrosine phosphorylation of nuclear-membrane protein emerin by Src, Abl and other kinases

Gap junction channel gating modulated through protein phosphorylation

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