title: Enterovirus 71 2C protein inhibits TNF-α-mediated activation of NF-κB by suppressing IκB kinase β phosphorylation.
abstract: Enterovirus 71 (EV71), a single, positive-stranded RNA virus, has been regarded as the most important neurotropic enterovirus after the eradication of the poliovirus. EV71 infection can cause hand, foot, and mouth disease or herpangina. Cytokine storm with elevated levels of proinflammatory and inflammatory cytokines, including TNF-α, has been proposed to explain the pathogenesis of EV71-induced disease. TNF-α-mediated NF-κB signaling pathway plays a key role in inflammatory response. We hypothesized that EV71 might also moderate host inflammation by interfering with this pathway. In this study, we tested this hypothesis and identified EV71 2C protein as an antagonist of TNF-α-mediated activation of NF-κB signaling pathway. Expression of 2C protein significantly reduced TNF-α-mediated NF-κB activation in 293T cells as measured by gene reporter and gel mobility shift assays. Furthermore, overexpression of TNFR-associated factor 2-, MEK kinase 1-, IκB kinase (IKK)α-, or IKKβ-induced NF-κB activation, but not constitutively active mutant of IKKβ (IKKβ SS/EE)-induced NF-κB activation, was inhibited by 2C protein. These data together suggested that the activation of IKKβ is most likely targeted by 2C; this notion was further strengthened by immunoblot detection of IKKβ phosphorylation and IκBα phosphorylation and degradation. Coimmunoprecipitation and colocalization of 2C and IKKβ expressed in mammalian cells provided compelling evidence that 2C interacts with IKKβ. Collectively, our data indicate that EV71 2C protein inhibits IKKβ activation and thus blocks NF-κB activation.
authors.affiliation: State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, 430071, China.
authors.forename: Zhenhua
authors.initials: Z
authors.lastname: Zheng
authors.forename: Hongxia
authors.initials: H
authors.lastname: Li
authors.forename: Zhenfeng
authors.initials: Z
authors.lastname: Zhang
authors.forename: Jin
authors.initials: J
authors.lastname: Meng
authors.forename: Da
authors.initials: D
authors.lastname: Mao
authors.forename: Bingke
authors.initials: B
authors.lastname: Bai
authors.forename: Baojing
authors.initials: B
authors.lastname: Lu
authors.forename: Panyong
authors.initials: P
authors.lastname: Mao
authors.forename: Qinxue
authors.initials: Q
authors.lastname: Hu
authors.forename: Hanzhong
authors.initials: H
authors.lastname: Wang
mesh_terms: D015153:Blotting, Western; D002352:Carrier Proteins / Q000276:immunology / Q000378:metabolism*; D024202:Electrophoretic Mobility Shift Assay; D029821:Enterovirus A, Human / Q000276:immunology; D004769:Enterovirus Infections / Q000276:immunology / Q000378:metabolism*; D004789:Enzyme Activation / Q000502:physiology*; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D051550:I-kappa B Kinase / Q000378:metabolism*; D047468:Immunoprecipitation; D008856:Microscopy, Fluorescence; D016328:NF-kappa B / Q000276:immunology / Q000378:metabolism*; D010766:Phosphorylation; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000502:physiology; D014162:Transfection; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology / Q000378:metabolism*; D017361:Viral Nonstructural Proteins / Q000276:immunology / Q000378:metabolism*
chemical_list: D002352:Carrier Proteins; D016328:NF-kappa B; D014409:Tumor Necrosis Factor-alpha; D017361:Viral Nonstructural Proteins; D051550:I-kappa B Kinase; C076023:2C protein, viral