Tyrosine phosphorylation of the Wiskott‐Aldrich Syndrome protein by Lyn and Btk is regulated by CDC42

Guinamard, Rodolphe; Aspenström, Pontus; Fougereau, Michel; Chavrier, Philippe; Guillemot, Jean‐Claude

doi:10.1016/s0014-5793(98)01016-3

Cited by 100 publications

(78 citation statements)

References 38 publications

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“…The WASP/suppressor of cAMP receptor family proteins hold potential for integrating such cytoskeletal and signaling responses. The more intensely studied WASP and N-WASP have well known effects on actin nucleation and in addition bind signaling proteins such as Src and Tec family tyrosine kinases and adapters such as Pro-Ser-Thr phosphatase-interacting protein, Grb2, and Nck (25)(26)(27)(28). Although WAVE1 has a clear role in the formation of Rac-induced ruffles and lamellipodia, its capacity to facilitate other signaling events is less clear.…”

Section: Discussionmentioning

confidence: 99%

Vascular Endothelial Growth Factor Causes Translocation of p47 to Membrane Ruffles through WAVE1

et al. 2003

Journal of Biological Chemistry

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Growth factors initiate cytoskeletal rearrangements tightly coordinated with nuclear signaling events. We hypothesized that the angiogenic growth factor, vascular endothelial growth factor (VEGF), may utilize oxidants that are site-directed to a complex critical to both cytoskeletal and mitogenic signaling. We identified the WASP-family verprolin homologous protein-1 (WAVE1) as a binding partner for the NADPH oxidase adapter p47 phox within membrane ruffles of VEGF-stimulated cells. Within 15 min of VEGF stimulation, p47 phox coprecipitated with WAVE1, with the ruffle and oxidase agonist Rac1, and with the Rac1 effector PAK1. VEGF also increased p47 phox phosphorylation, oxidant production, and ruffle formation, all of which were dependent upon PAK1 kinase activity. The antioxidant Mn (III) tetrakis(4-benzoic acid) porphyrin and ectopic expression of either the p47-binding WAVE1 domain or the WAVE1-binding p47 phox domain decreased VEGF-induced ruffling, whereas the active mutant p4-(S303D, S304D,S328D) stimulated oxidant production and formation of circular dorsal ruffles. Both kinase-dead PAK1-(K298A) and Mn (III) tetrakis(4-benzoic acid) porphyrin decreased c-Jun N-terminal kinase (JNK) activation by VEGF, whereas dominant-negative JNK did not block ruffle formation, suggesting a bifurcation of mitogenic and cytoskeletal signaling events at or distal to the oxidase but proximal to JNK. Thus, WAVE1 may act as a scaffold to recruit the NADPH oxidase to a complex involved with both cytoskeletal regulation and downstream JNK activation.

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Section: Discussionmentioning

confidence: 99%

Vascular Endothelial Growth Factor Causes Translocation of p47 to Membrane Ruffles through WAVE1

et al. 2003

Journal of Biological Chemistry

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“…FAK Phosphorylation of N-WASP at Tyr 256 -A number of tyrosine kinases have been shown to associate with WASP family proteins and phosphorylate them (29,(33)(34)(35)(36). To determine whether N-WASP is a substrate for FAK, we first tested whether purified FAK can phosphorylate N-WASP in vitro.…”

Section: Association Of Fak With N-wasp-mentioning

confidence: 99%

“…4A shows that mutation of Tyr 256 to Phe (Y256F) significantly reduced N-WASP phosphorylation by FAK, suggesting that Tyr 256 of N-WASP is the major FAK phosphorylation site. Recent structural and biochemical studies indicated that Cdc42 binding to WASP can increase the accessibility of Tyr 291 for phosphorylation by the tyrosine kinases, Lyn and Btk (35,38). We therefore tested whether phosphorylation of Tyr 256 of N-WASP by FAK is dependent on its activation state.…”

Section: Association Of Fak With N-wasp-mentioning

confidence: 99%

Focal Adhesion Kinase Regulation of N-WASP Subcellular Localization and Function

Suetsugu

Cooper

et al. 2004

Journal of Biological Chemistry

169

143

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N-WASP is a member of the WASP family of proteins that regulate actin cytoskeleton remodeling. FAK is a cytoplasmic tyrosine kinase implicated in integrin signaling during cell migration. Here we identify a direct interaction between N-WASP and FAK and show that N-WASP is phosphorylated by FAK at a conserved tyrosine residue, Tyr 256 . We found that phosphorylation of Tyr 256 affected N-WASP nuclear localization, suggesting that phosphorylation of N-WASP by FAK may regulate its activity in vivo by altering its subcellular localization. We also showed that the nuclear localization of N-WASP is dependent on its being in the open conformation either after its activation by Cdc42 or the truncation of the C-terminal VCA domain. Phosphorylation of Tyr 256 of N-WASP could reduce its interaction with nuclear importin NPI-1, which might be responsible for its decreased nuclear localization. Lastly, we show that phosphorylation of Tyr 256 plays an important role in promoting cell migration. Together, these results suggest a novel regulatory mechanism of N-WASP by tyrosine phosphorylation and subcellular localization and its potential role in the regulation of cell migration.

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“…This defect in phagocytosis was also observed in preliminary flow cytometric experiments (data not shown). Defective phagocytosis in Btk-deficient macrophages is interesting in light of the reported connections between Btk, Wiskott-Aldrich syndrome protein, Cdc42, and actin polymerization (37)(38)(39).…”

Section: Btk-deficient Macrophages Show Poor Microbicidal Functionsmentioning

confidence: 99%

Macrophage Effector Functions Controlled by Bruton’s Tyrosine Kinase Are More Crucial Than the Cytokine Balance of T Cell Responses for Microfilarial Clearance

Mukhopadhyay

Mohanty

Mangla

et al. 2002

The Journal of Immunology

100

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Macrophages from X-linked immunodeficient (xid) mice lacking functional Bruton’s tyrosine kinase (Btk) show poor NO induction and enhanced IL-12 induction, and contribute to delayed clearance of injected microfilaria (mf) in vivo. We now show that Btk is involved in other macrophage effector functions, such as bactericidal activity and secretion of proinflammatory cytokines (TNF-α, IL-1β), but not the T cell-directed cytokine IL-12. Induction of some transcriptional regulators of the NF-κB family, crucial for the expression of proinflammatory cytokines, is also poor in Btk-deficient macrophages. Thus, Btk appears to be involved in signaling for inducible effector functions, but not APC functions, in macrophages. Furthermore, adoptive transfer of T cells from mf-infected xid or wild-type mice did not alter the course of mf clearance in recipients, mf clearance was unaltered in IFN-γ-deficient mice, and improved mf clearance was seen only if greater inducibility of IL-12 was accompanied by greater NO secretion from macrophages, as seen in Ityr C.D2 mice as compared with congenic Itys BALB/c mice. Thus, delayed mf clearance in xid mice was correlated not with the high IL-12/Th1 phenotype but with low NO induction levels. Also, xid macrophages showed poor toxicity to mf in vitro as compared with wild-type macrophages. Inhibition of NO production blocked this mf cytotoxicity, and an NF-κB inhibitor blocked both NO induction and mf cytotoxicity. Thus, Btk is involved in inducing many macrophage effector functions, and delayed mf clearance seen in Btk-deficient xid mice is due to poor NO induction in macrophages, resulting in compromised microfilarial toxicity.

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Tyrosine phosphorylation of the Wiskott‐Aldrich Syndrome protein by Lyn and Btk is regulated by CDC42

Cited by 100 publications

References 38 publications

Vascular Endothelial Growth Factor Causes Translocation of p47 to Membrane Ruffles through WAVE1

Vascular Endothelial Growth Factor Causes Translocation of p47 to Membrane Ruffles through WAVE1

Focal Adhesion Kinase Regulation of N-WASP Subcellular Localization and Function

Macrophage Effector Functions Controlled by Bruton’s Tyrosine Kinase Are More Crucial Than the Cytokine Balance of T Cell Responses for Microfilarial Clearance

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