Ctenopharyngodon idella IKKβ interacts with PKR and IκBα
Ctenopharyngodon idella IKKβ interacts with PKR and IκBα
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草鱼 IKK beta 与 PKR 和 I kappa B alpha 相互作用
DOI：
10.1093/abbs/gmx065
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发表时间：
2017
期刊：
影响因子：
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通讯作者：
Chengyu Hu
中科院分区：
文献类型：
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作者：
Haizhou Wang;Qun Xu;Yousheng HU;Youlin Zhu;Chengyu Hu
核因子 kappa-B 激酶 β 抑制剂 (IKKβ) 是 IKK 复合物的一个亚基。它可以响应广泛的刺激，通过磷酸化 IκB 来激活 NF-κB 通路。本研究克隆了来自草鱼（Ctenopharyngodon idella；KT282114）的IKKβ基因，并通过同源克隆和cDNA末端快速扩增（RACE）技术进行鉴定。完整的CiIKKβ cDNA长度为3428 bp，最长开放阅读框（ORF）为2337 bp，编码778个氨基酸的多肽。推导的.CiIKKβ氨基酸序列具有与哺乳动物相似的结构域分布。例如，CiIKKβ由N端的丝氨酸/苏氨酸激酶结构域、中间的碱性区域亮氨酸拉链(BRLZ)结构域、同源盒相关的亮氨酸拉链(HALZ)结构域和C端的IKKβNEMO(NF-κB必需调节剂)结合结构域组成。系统发育树分析还表明，CiIKKβ 与斑马鱼 IKKβ (DrIKKβ) 高度同源，并且与哺乳动物和两栖动物的对应物明显不同。 CiIKKβ在草鱼的肝、肠、肾、鳃、脾、心脏和脑组织中普遍存在，并且在Poly I:C和紫外线灭活草鱼出血病毒的刺激下，CIK细胞中的表达显着上调。为了研究鱼类NF-κB通路的激活机制以及.CiIKKβ在该通路中的作用，我们通过免疫共沉淀和GST-pull down实验探讨了蛋白激酶R（PKR）与IKKβ以及IKKβ与IκBα的蛋白质相互作用。每对之间的相互作用得到证实。结果表明CiIKKβ可能是鱼类NF-κB通路激活的主要成员。
Inhibitor of nuclear factor kappa-B kinase β (IKKβ) is a subunit of the IKK complex. It can activate.the NF-κB pathway through phosphorylating IκB in response to a wide range of stimuli. In the.present study, an IKKβ gene from grass carp (Ctenopharyngodon idella; KT282114) was cloned.and identified by homologous cloning and rapid-amplification of cDNA ends (RACE) technique..The complete CiIKKβ cDNA is 3428 bp in length, with the longest open reading frame (ORF) of.2337 bp encoding a polypeptide of 778 amino acids. The deduced amino acid sequence of.CiIKKβ has similar domain distribution to those of mammalian. For example, CiIKKβ consists of.a serine/threonine kinase domain at the N-terminal, a basic region leucin zipper (BRLZ) domain.in the middle, a homeobox associated leucin zipper (HALZ) domain and an IKKβ NEMO (NF-κB.essential modulator) binding domain at the C-terminal. Phylogenetic tree analysis also showed.that CiIKKβ is highly homologous to zebrafish IKKβ (DrIKKβ) and clearly distinct from the mammalian.and amphibian counterparts. The expression of CiIKKβ was ubiquitously found in the.liver, intestine, kidney, gill, spleen, heart, and brain tissues of grass carp and significantly.up-regulated in CIK cells under the stimulation with Poly I:C and UV-inactivated grass carp hemorrhagic.virus. To investigate the activation mechanism of NF-κB pathway in fish and the role of.CiIKKβ in the pathway, we explored the protein interactions of protein kinase R (PKR) with IKKβ.and IKKβ with IκBα by co-immunoprecipitation and GST-pull down assays. The interaction.between each pair was confirmed. The results suggest that CiIKKβ may be a primarymember in.the activation of NF-κB pathway in fish.