c-Src-dependent Tyrosine Phosphorylation of IKKβ Is Involved in Tumor Necrosis Factor-α-induced Intercellular Adhesion Molecule-1 Expression

Huang, Wei-Chien; Chen, Jun Jie; Chen, Ching-Chow

doi:10.1074/jbc.m208521200

Cited by 120 publications

(109 citation statements)

References 45 publications

(42 reference statements)

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“…The role of c-Src in TNF-induced NF-B activation has been demonstrated (21,23). It has been shown that c-Src causes the tyrosine phosphorylation of I B kinase-␤ needed for TNF-induced NF-B activation (23).…”

Section: Discussionmentioning

confidence: 99%

“…Proline-rich tyrosine kinase (Pyk2) 3 (18,19) and p53/Lyn p56 (20) have roles in TNF-induced neutrophil activation; Etk/Bmx in TNF-induced endothelial cell migration and angiogenesis; c-Src in TNF-induced NF-B activation in the marrow macrophages (21,22); and TNF in regulation of gap junctions in airway epithelial cells (23). Pyk2 has also been implicated in TNF-induced JNK activation (24).…”

Section: Tnf Activates Syk Proteinmentioning

confidence: 99%

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TNF Activates Syk Protein Tyrosine Kinase Leading to TNF-Induced MAPK Activation, NF-κB Activation, and Apoptosis

Takada

Aggarwal

2004

The Journal of Immunology

108

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Spleen tyrosine kinase (Syk), a nonreceptor protein kinase initially found to be expressed only in hemopoietic cells, has now been shown to be expressed in nonhemopoietic cells and to mediate signaling of various cytokines. Whether Syk plays any role in TNF signaling was investigated. Treatment of Jurkat T cells with TNF activated Syk kinase but not ZAP70, another member of Syk kinase family, and the optimum activation occurred at 10 s and with 1 nM TNF. TNF also activated Syk in myeloid and epithelial cells. TNF-induced Syk activation was abolished by piceatannol (Syk-selective inhibitor), which led to the suppression of TNF-induced activation of c- JNK, p38 MAPK, and p44/p42 MAPK. Jurkat cells that did not express Syk (JCaM1, JCaM1/lck) showed lack of TNF-induced Syk, JNK, p38 MAPK, and p44/p42 MAPK activation, as well as TNF-induced IκBα phosphorylation, IκBα degradation, and NF-κB activation. TNF-induced NF-κB activation was enhanced by overexpression of Syk by Syk-cDNA and suppressed when Syk expression was down-regulated by expression of Syk-small interfering RNA (siRNA-Syk). The apoptotic effects of TNF were reduced by up-regulation of NF-κB by Syk-cDNA, and enhanced by down-regulation of NF-κB by siRNA-Syk. Immunoprecipitation of cells with Syk Abs showed TNF-dependent association of Syk with both TNFR1 and TNFR2; this association was enhanced by up-regulation of Syk expression with Syk-cDNA and suppressed by down-regulation of Syk using siRNA-Syk. Overall, our results demonstrate that Syk activation plays an essential role in TNF-induced activation of JNK, p38 MAPK, p44/p42 MAPK, NF-κB, and apoptosis.

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Section: Discussionmentioning

confidence: 99%

Section: Tnf Activates Syk Proteinmentioning

confidence: 99%

TNF Activates Syk Protein Tyrosine Kinase Leading to TNF-Induced MAPK Activation, NF-κB Activation, and Apoptosis

Takada

Aggarwal

2004

The Journal of Immunology

108

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“…Our previous data showed that in contrast to other members of Src family, c-Src and Lyn were abundantly expressed in NCI-H292 cell and another human alveolar epithelial A549 cell (26). c-Src and Lyn in NCI-H292 cells were therefore isolated by immunoprecipitation using anti-c-Src or anti-Lyn Ab, and their in vitro kinase activity was measured using enolase as substrate.…”

Section: Induction Of C-src and Lyn Activation By Tnf-␣ And Tpa And mentioning

confidence: 99%

“…Tyrosine (Y) 199, tyrosine 188, or both sites in the human IKK␤ cloned in the pcDNA3.1 vector, or in the human GST-IKK␤ (132-206) cloned in the pGEX vector was substituted with phenylalanine (F) by changing the triplet from TAC to TTC. The mutated primers used were primer 1 (5Ј-CGAGGGTTGCCATCATGACTCTGAAGCCAGGCA-3Ј) and primer 2 (3Ј-GCTCCCAACGGTAGTACTGAGACTTCGGTCCGT-5Ј) for c-Src (K295 M) mutation, primer 3 (5Ј-GGGGACCCTGCAGTTC CTGGCCCCAGAGC-3Ј), primer 4 (3Ј-CCCCTGGGACGTCAAGGACCG GGGTCTCG-5Ј) for IKK␤ (Y188F) mutation, primer 5 (5Ј-GGAGCAG CAGAAGTTCACAGTGACCGTCG-3Ј), and primer 6 (3Ј-CCTCGTCGT CTTCAAGTGTCACTGGCAGC-5Ј) for IKK␤ (Y199F) mutation, as described previously (26).…”

Section: Site-directed Mutagenesismentioning

confidence: 99%

“…8C). Assuming that Tyr 188 and/or Tyr 199 of IKK␤ were the targets for c-Src phosphorylation after TNF-␣ or TPA stimulation, we used site-directed mutagenesis to generate the dominant-negative tyrosine mutants, IKK␤ (Y188F), IKK␤ (Y199F), and IKK␤ (Y188F, Y199F) (26). Overexpression of these mutants attenuated the TNF-␣-or TPA-induced COX-2 promoter activity (Fig.…”

Section: Inhibitory Effect Of the Dominant-negative Mutants Ikk␤ (Y1mentioning

confidence: 99%

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Tyrosine Phosphorylation of I-κB Kinase α/β by Protein Kinase C-Dependent c-Src Activation Is Involved in TNF-α-Induced Cyclooxygenase-2 Expression

Huang

Chen

Inoue

et al. 2003

The Journal of Immunology

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116

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The signaling pathway involved in TNF-α-induced cyclooxygenase-2 (COX-2) expression was further studied in human NCI-H292 epithelial cells. A protein kinase C (PKC) inhibitor (staurosporine), tyrosine kinase inhibitors (genistein and herbimycin A), or a Src kinase inhibitor (PP2) attenuated TNF-α- or 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced COX-2 promoter activity. TNF-α- or TPA-induced I-κB kinase (IKK) activation was also blocked by these inhibitors, which reversed I-κBα degradation. Activation of c-Src and Lyn kinases, two Src family members, was inhibited by the PKC, tyrosine kinase, or Src kinase inhibitors. The dominant-negative c-Src (KM) mutant inhibited induction of COX-2 promoter activity by TNF-α or TPA. Overexpression of the constitutively active PKCα (PKCα A/E) or wild-type c-Src plasmids induced COX-2 promoter activity, and these effects were inhibited by the dominant-negative c-Src (KM), NF-κB-inducing kinase (NIK) (KA), or IKKβ (KM) mutant. The dominant-negative PKCα (K/R) or c-Src (KM) mutant failed to block induction of COX-2 promoter activity caused by wild-type NIK overexpression. In coimmunoprecipitation experiments, IKKα/β was found to be associated with c-Src and to be phosphorylated on its tyrosine residues after TNF-α or TPA treatment. Two tyrosine residues, Tyr188 and Tyr199, near the activation loop of IKKβ, were identified to be crucial for NF-κB activation. Substitution of these residues with phenylalanines attenuated COX-2 promoter activity and c-Src-dependent phosphorylation of IKKβ induced by TNF-α or TPA. These data suggest that, in addition to activating NIK, TNF-α also activates PKC-dependent c-Src. These two pathways cross-link between c-Src and NIK and converge at IKKα/β, and go on to activate NF-κB, via serine phosphorylation and degradation of IκB-α, and, finally, to initiate COX-2 expression.

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Src kinase Lyn is crucial for Pseudomonas aeruginosa internalization into lung cells

et al. 2006

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Lyn is an important B cell signaling kinase of the Src tyrosine kinase family with a broad range of functions from cytoskeletal changes to induction of apoptosis. However, the role of Lyn in infectious diseases is not clear. Here, we demonstrate that Lyn activation by phosphorylation significantly impacted invasion of an alveolar epithelial cell line, primary lung cells, and rat lungs by Pseudomonas aeruginosa (PA), a common opportunistic lung pathogen affecting individuals with deficient lung immunity. Our results indicate that activation of Lyn and its interaction with rafts and TLR2, played an important role in the initial stages of PA interaction with host cells. The role of Lyn was further evaluated using the pharmacologic Src-specific inhibitor PP2, a dominant negative mutant, and finally confirmed with Lyn-deficient (Lyn -/-) bone marrowderived mast cells. Inhibition of Lyn's function by above approaches prevented PA internalization. Moreover, blocking of Lyn also affected downstream events: induction of inflammatory cytokines and apoptosis. This report brings out a new role of Lyn in infectious diseases and indicates potential new targets for prevention and treatment of infections.

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c-Src-dependent Tyrosine Phosphorylation of IKKβ Is Involved in Tumor Necrosis Factor-α-induced Intercellular Adhesion Molecule-1 Expression

Cited by 120 publications

References 45 publications

TNF Activates Syk Protein Tyrosine Kinase Leading to TNF-Induced MAPK Activation, NF-κB Activation, and Apoptosis

TNF Activates Syk Protein Tyrosine Kinase Leading to TNF-Induced MAPK Activation, NF-κB Activation, and Apoptosis

Tyrosine Phosphorylation of I-κB Kinase α/β by Protein Kinase C-Dependent c-Src Activation Is Involved in TNF-α-Induced Cyclooxygenase-2 Expression

Src kinase Lyn is crucial for Pseudomonas aeruginosa internalization into lung cells

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