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凡纳滨对虾肿瘤坏死因子受体相关因子 6(TRAF6)对溶藻弧菌和白斑综合征病毒(WSSV)感染的反应及激活抗菌肽基因。

Litopenaeus vannamei tumor necrosis factor receptor-associated factor 6 (TRAF6) responds to Vibrio alginolyticus and white spot syndrome virus (WSSV) infection and activates antimicrobial peptide genes.

机构信息

State Key Laboratory of Biocontrol/MOE Key Laboratory of Aquatic Product Safety, School of Life Sciences, Sun Yat-sen University, 135 Xingang Road West, Guangzhou 510275, People's Republic of China.

出版信息

Dev Comp Immunol. 2011 Jan;35(1):105-14. doi: 10.1016/j.dci.2010.08.013. Epub 2010 Sep 17.

DOI:10.1016/j.dci.2010.08.013
PMID:20816892
Abstract

Tumor necrosis factor receptor (TNFR)-associated factor 6 (TRAF6) is a key signaling adaptor protein not only for the TNFR superfamily but also for the Interleukin-1 receptor/Toll-like receptor (IL-1/TLR) superfamily. To investigate TRAF6 function in invertebrate innate immune responses, Litopenaeus vannamei TRAF6 (LvTRAF6) was identified and characterized. The full-length cDNA of LvTRAF6 is 2823bp long, with an open reading frame (ORF) encoding a putative protein of 594 amino acids, including a RING-type Zinc finger, two TRAF-type Zinc fingers, a coiled-coil region, and a meprin and TRAF homology (MATH) domain. The overall amino acid sequence identity between LvTRAF6 and other known TRAF6s is 22.2-33.3%. Dual luciferase reporter assays in Drosophila S2 cells revealed that LvTRAF6 could activate the promoters of antimicrobial peptide genes (AMPs), including Drosophila Attacin A and Drosomycin, and shrimp Penaeidins. Real-time quantitative PCR (qPCR) indicated that LvTRAF6 was constitutively expressed in various tissues of L. vannamei. After Vibrio alginolyticus and white spot syndrome virus (WSSV) challenge, LvTRAF6 was down-regulated, though with different expression patterns in the intestine compared to other tissues. After WSSV challenge, LvTRAF6 was up-regulated 2.7- and 2.3-fold over the control at 3h in gills and hepatopancreas, respectively. These results indicated that LvTRAF6 may play a crucial role in antibacterial and antiviral responses via regulation of AMP gene expression.

摘要

肿瘤坏死因子受体(TNFR)相关因子 6(TRAF6)不仅是 TNFR 超家族,也是白细胞介素-1 受体/ Toll 样受体(IL-1/TLR)超家族的关键信号转导衔接蛋白。为了研究 TRAF6 在无脊椎动物先天免疫反应中的功能,鉴定并表征了凡纳滨对虾 TRAF6(LvTRAF6)。LvTRAF6 的全长 cDNA 长 2823bp,包含一个开放阅读框(ORF),编码一个 594 个氨基酸的假定蛋白,包括一个 RING 型锌指、两个 TRAF 型锌指、一个卷曲螺旋区和一个髓过氧化物酶和 TRAF 同源(MATH)结构域。LvTRAF6 与其他已知 TRAF6 之间的整体氨基酸序列同一性为 22.2-33.3%。在果蝇 S2 细胞中的双荧光素酶报告基因检测表明,LvTRAF6 可以激活抗菌肽基因(AMPs)的启动子,包括果蝇 Attacin A 和 Drosomycin 以及虾的 Penaeidins。实时定量 PCR(qPCR)表明,LvTRAF6 在凡纳滨对虾的各种组织中均呈组成型表达。在受到副溶血弧菌和白斑综合征病毒(WSSV)的攻击后,LvTRAF6 的表达下调,尽管在肠组织中的表达模式与其他组织不同。在受到 WSSV 攻击后,LvTRAF6 在鳃和肝胰腺中的表达分别比对照组上调了 2.7 倍和 2.3 倍。这些结果表明,LvTRAF6 可能通过调节 AMP 基因的表达在抗菌和抗病毒反应中发挥关键作用。

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