Respiratory syncytial virus influences NF-κB-dependent gene expression through a novel pathway involving MAP3K14/NIK expression and nuclear complex formation with NF-κB2
Respiratory syncytial virus influences NF-κB-dependent gene expression through a novel pathway involving MAP3K14/NIK expression and nuclear complex formation with NF-κB2
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DOI：
10.1128/jvi.79.14.8948-8959.2005
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发表时间：
2005-07-01
影响因子：
5.4
通讯作者：
Brasier, AR
中科院分区：
文献类型：
--
作者：
Choudhary, S;Boldogh, S;Brasier, AR
呼吸道合胞病毒（RSV）是副粘病毒科RNA病毒的一员，是儿童流行性呼吸道感染的主要原因。在儿童中，RSV主要在气道粘膜中复制，这一过程改变了上皮细胞趋化因子的表达，从而诱导气道炎症。我们研究了丝裂原活化蛋白激酶激酶激酶14/NF-kappa b诱导激酶（NIK）在肺泡样A549细胞NF-kappa b依赖性基因激活中的作用。RSV感染诱导NIK mRNA和蛋白表达的时间依赖性增加，在病毒暴露后12至24小时达到峰值。免疫沉淀激酶试验表明，NIK激酶活性被更快地激活（在RSV吸附后6小时内），这与内源性类似于50 kda的NF-kappa B2底物有关。由于NIK与IKK α结合介导100 kda NF-kappa B2前体转化为52-kDa DNA结合异构体（“p52”），RSV对IKKa和NF-kappa B2形成NIK复合物的影响通过共免疫沉淀法确定。我们发现NIK、IKKa和100 kDa和52-kDa NF-kappa B2在暴露于RSV 15小时后形成强烈的复合物，有时会在NIK激酶激活后形成。Western免疫印迹和微亲和DNA拉下实验显示NF-kappa b2 - re1b复合物的核易位和DNA结合平行增加。有趣的是，我们观察到NIK也会瞬时易位到与52-kDa NF-kappa B2复合物的细胞核中。小干扰rna介导的NIK“敲低”阻断了RSV诱导的52-kDa NF-kappa B2加工，干扰了NF-kappa B依赖性基因亚群的早期激活，表明该激活途径在基因组NF-kappa B对RSV的应答中具有重要意义。总之，这些数据表明，RSV感染在更有效的规范途径激活之前，迅速激活非规范NF-kappa B激活途径。这似乎是通过一种新的机制，包括诱导NIK激酶活性、表达和IKK α和加工NF-kappa B2三元复合物的核易位。
A member of the Paramyxoviridae family of RNA viruses, respiratory syncytial virus (RSV), is a leading cause of epidemic respiratory tract infection in children. In children, RSV primarily replicates in the airway mucosa, a process that alters epithelial cell chemokine expression, thereby inducing airway inflammation. We investigated the role of the mitogen-activated protein kinase kinase kinase 14/NF-kappa B-inducing kinase (NIK) in the activation of NF-kappa B-dependent genes in alveolus-like A549 cells. RSV infection induces a time dependent increase of NIK mRNA and protein expression that peaks 12 to 24 h after viral exposure. Immunoprecipitation kinase assays indicate that NIK kinase activity is activated even more rapidly (within 6 h of RSV adsorption) associated with an endogenous similar to 50-kDa NF-kappa B2 substrate. Because NIK associates with IKK alpha to mediate processing of the 100-kDa NF-kappa B2 precursor into its 52-kDa DNA binding isoform ("p52"), the effects of RSV on NIK complex formation with IKKa and NF-kappa B2 were determined by coimmunoprecipitation assay. We find that NIK, IKKa, and both 100 kDa- and 52-kDa NF-kappa B2 isoforms strongly complex 15 h after exposure to RSV at times subsequent to NIK kinase activation. Western immunoblot and microaffinity DNA pull-down assays showed a parallel increase in nuclear translocation and DNA binding of the NF-kappa B2-Re1 B complex. Interestingly, we make the novel observations that NIK also transiently translocates into the nucleus complexed with 52-kDa NF-kappa B2. Small interfering RNA-mediated NIK "knock-down" blocked RSV-inducible 52-kDa NF-kappa B2 processing and interfered with the early activation of a subset of NF-kappa B-dependent genes, indicating the importance of this activation pathway in the genomic NF-kappa B response to RSV. Together, these data indicate that RSV infection rapidly activates the noncanonical NF-kappa B activation pathway prior to the more potent canonical pathway activation. This appears to be through a novel mechanism involving induction of NIK kinase activity, expression, and nuclear translocation of a ternary complex with IKK alpha and processed NF-kappa B2.