Qu Yixiao, Zhou Man, Peng Linzhi, Li Jun, Yan Jun, Yang Peilin, Feng Hao
Key Laboratory of Protein Chemistry and Developmental Biology of Ministry of Education of China, College of Life Science, Hunan Normal University, Changsha, 410081, China.
Key Laboratory of Protein Chemistry and Developmental Biology of Ministry of Education of China, College of Life Science, Hunan Normal University, Changsha, 410081, China.
Fish Shellfish Immunol. 2015 Nov;47(1):122-9. doi: 10.1016/j.fsi.2015.08.033. Epub 2015 Aug 30.
IKKε is an IκB kinase functioning in NF-κB signal pathway in the innate immune system of higher vertebrates. To exploit the function of IKKε of black carp (bcIKKε) in its antiviral innate immunity, the IKKε gene has been cloned from the RNA isolated from the spleen of black carp. The full-length cDNA of bcIKKε is 2537 bp, which encodes the peptide of 723 amino acids. bcIKKε contains a S-Tkc domain, a PKc domain and a UBL-TBK1-like domain and bcIKKε shares the highest amino acid sequence similarity with that of grass carp. bcIKKε was constitutively transcribed in the selected tissues of black carp including gill, kidney, heart, intestine, liver, muscle, skin and spleen; and the mRNA level of bcIKKε in these tissues varied right after SVCV or GCRV infection. bcIKKε had been well expressed in HEK293T cells and western blot assay determined that this fish kinase was around 80 KDa. The immunofluorescence assay of both NH3T3 cells and EPC cells demonstrated that bcIKKε was located in the cytosolic part of the cell. Report assay result showed that overexpression of bcIKKε in EPC cells activated the expression of both zebrafish IFN and EPC IFN. All our data suggest that bcIKKε is a novel fish kinase functioning in the innate antiviral immune response of black carp.
IKKε是一种在高等脊椎动物先天免疫系统的NF-κB信号通路中发挥作用的IκB激酶。为了探究草鱼IKKε（bcIKKε）在其抗病毒先天免疫中的功能，已从草鱼脾脏分离的RNA中克隆出IKKε基因。bcIKKε的全长cDNA为2537 bp，编码723个氨基酸的肽段。bcIKKε包含一个S-Tkc结构域、一个PKc结构域和一个UBL-TBK1样结构域，且bcIKKε与草鱼的氨基酸序列相似度最高。bcIKKε在草鱼的鳃、肾、心脏、肠道、肝脏、肌肉、皮肤和脾脏等选定组织中组成性转录；在感染鲤春病毒血症病毒（SVCV）或草鱼呼肠孤病毒（GCRV）后，这些组织中bcIKKε的mRNA水平立即发生变化。bcIKKε在人胚肾293T细胞（HEK293T细胞）中表达良好，蛋白质免疫印迹分析确定这种鱼类激酶约为80 kDa。对小鼠成纤维细胞（NH3T3细胞）和草鱼肾细胞（EPC细胞）的免疫荧光分析表明，bcIKKε位于细胞的胞质部分。报告基因检测结果显示，bcIKKε在EPC细胞中的过表达激活了斑马鱼干扰素（IFN）和EPC干扰素的表达。我们所有的数据表明，bcIKKε是一种在草鱼先天抗病毒免疫反应中发挥作用的新型鱼类激酶。
