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凡纳滨对虾TAB2基因的鉴定与功能表征

Identification and functional characterization of the TAB2 gene from Litopenaeus vannamei.

作者信息

Wang Sheng, Li Haoyang, Qian Zhe, Song Xuan, Zhang Zijian, Zuo Hongliang, Xu Xiaopeng, Weng Shaoping, He Jianguo, Li Chaozheng

机构信息

MOE Key Laboratory of Aquatic Product Safety/State Key Laboratory for Biocontrol, School of Life Sciences, Sun Yat-sen University, Guangzhou, PR China; Institute of Aquatic Economic Animals and Guangdong Provice Key Laboratory for Aquatic Economic Animals, Sun Yat-sen University, Guangzhou, PR China; Guangdong Provincial Key Laboratory of Marine Resources and Coastal Engineering, Sun Yat-sen University, Guangzhou, PR China.

MOE Key Laboratory of Aquatic Product Safety/State Key Laboratory for Biocontrol, School of Life Sciences, Sun Yat-sen University, Guangzhou, PR China.

出版信息

Fish Shellfish Immunol. 2015 Oct;46(2):206-16. doi: 10.1016/j.fsi.2015.06.024. Epub 2015 Jun 20.

Abstract

In Drosophila, TAB2, an important intermediate in the IMD signaling pathway, plays critical roles in the innate immune response in response to bacterial and viral infection. However, the role of TAB-related proteins in the immune response of shrimp has not yet been established. Here, we reported the identification of a TAB2-like gene in Litopenaeus vannamei designated as LvTAB2. The full-length cDNA of LvTAB2 was 2160 bp with an open reading frame of 1827 bp, which encoded a putative protein of 608 amino acids including a ubiquitin binding domain (CUE) at the N-terminal and a Zinc Finger domain (ZnF) at the C-terminus. Real-time RT-PCR analysis showed that LvTAB2 was expressed in all tested tissues and the expression levels of LvTAB2 in gills and hemocytes were positively induced in response to LPS, Vibrio parahemolyticus and White Spot Syndrome Virus (WSSV) challenges. Dual luciferase reporter assays demonstrated that LvTAB2 was able to induce the expression of antimicrobial peptide (AMP) genes, including Drosophila Attacin A and shrimp Penaeidins. Interestingly, over-expression of LvTAB2 could up-regulate the promoter activities of L. vannamei Vago1, Vago3 and Vago4 genes in S2 cells. To our knowledge, it was the first report that TAB2 participated in innate immune signaling to regulate the expression of Vago genes in invertebrates. Moreover, RNAi-mediated knockdown of LvTAB2 enhanced sensitivity of L. vannamei to Vibrio parahaemolyticus infection and caused elevated virus loads after WSSV infection. We suggested that the LvTAB2 may play important roles in the shrimp innate immunity.

摘要

在果蝇中,TAB2是IMD信号通路中的一个重要中间体,在应对细菌和病毒感染的先天免疫反应中发挥关键作用。然而,TAB相关蛋白在虾免疫反应中的作用尚未明确。在此,我们报道了在凡纳滨对虾中鉴定出一个类似TAB2的基因,命名为LvTAB2。LvTAB2的全长cDNA为2160 bp,开放阅读框为1827 bp,编码一个推定的含有608个氨基酸的蛋白质,该蛋白质在N端包含一个泛素结合结构域(CUE),在C端包含一个锌指结构域(ZnF)。实时RT-PCR分析表明,LvTAB2在所有测试组织中均有表达,并且在受到脂多糖、副溶血性弧菌和白斑综合征病毒(WSSV)刺激后,鳃和血细胞中LvTAB2的表达水平呈阳性诱导。双荧光素酶报告基因检测表明,LvTAB2能够诱导抗菌肽(AMP)基因的表达,包括果蝇防御素A和虾对虾素。有趣的是,LvTAB2的过表达能够上调凡纳滨对虾Vago1、Vago3和Vago4基因在S2细胞中的启动子活性。据我们所知,这是首次报道TAB2参与无脊椎动物的先天免疫信号传导以调节Vago基因的表达。此外,RNA干扰介导的LvTAB2敲低增强了凡纳滨对虾对副溶血性弧菌感染的敏感性,并在WSSV感染后导致病毒载量升高。我们认为LvTAB2可能在虾的先天免疫中发挥重要作用。

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