Shrimp MyD88 responsive to bacteria and white spot syndrome virus.
Shrimp MyD88 responsive to bacteria and white spot syndrome virus.
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DOI：
10.1016/j.fsi.2012.11.034
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发表时间：
2013-02
影响因子：
4.7
通讯作者：
R. Wen;Fuhua Li;Zheng Sun;Shihao Li;J. Xiang
中科院分区：
文献类型：
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作者：
R. Wen;Fuhua Li;Zheng Sun;Shihao Li;J. Xiang
髓样分化因子88 （MyD88）是连接toll样受体（TLR）成员与下游组分激活相关信号通路的重要适配蛋白。本研究从中国对虾Fenneropenaeus chinensis中克隆了一个MyD88同源基因FcMyD88。FcMyD88的ORF全长1434 bp，编码一个477个氨基酸的多肽，包含一个死亡结构域（DD）和一个典型的TLR和白细胞介素-1受体（IL-1R）相关结构域（TIR）。同源性分析显示，预测的FcMyD88氨基酸（aa）序列与先前报道的多种myd88具有高度相似性。采用real-time PCR和western blotting分别分析了FcMyD88在注射了革兰氏阴性菌——鳗弧菌（G−）、革兰氏阳性菌——溶球微球菌（G +）和白色综合征斑状病毒（WSSV）的对虾头胸腔中转录和蛋白水平的时间依赖性表达模式。FcMyD88 mRNA的表达水平在鳗弧菌和溶血弧菌刺激后1小时(h)、12小时和24小时均显著上调。FcMyD88蛋白的表达量在注射灭活鳗鲡弧菌12 h （hpi）时上调2倍，而在此期间注射溶菌弧菌后表达量没有变化。注射WSSV后，FcMyD88 mRNA的表达水平保持相对恒定，而FcMyD88蛋白在12和24 hpi时显著上调。这些结果表明myd88依赖的信号通路可能参与细菌和WSSV感染的防御。
The myeloid differentiation factor 88 (MyD88) is an important adapter protein which links members of the toll-like receptor (TLR) to the downstream components to activate related signaling pathways. In the present study, a MyD88 homolog (FcMyD88) was cloned from penaeid shrimp Fenneropenaeus chinensis. The ORF of FcMyD88 consisted of 1434 bp encoding a polypeptide of 477 amino acids which contains a death domain (DD) and a typical TLR and interleukin-1 receptor (IL-1R)-related (TIR) domain. Homology analysis revealed that the predicted amino acid (aa) sequence of FcMyD88 shared high similarities with a variety of previously reported MyD88s. The time-dependent expression patterns of FcMyD88 in cephalothoraxes of shrimp injected with Vibrio anguillarum (Gram-negative bacteria, G−), Micrococcus lysodeikticu (Gram-positive bacteria, G+) and white syndrome spot virus (WSSV) were analyzed at transcription and protein level by real-time PCR and western blotting, respectively. The expression level of FcMyD88 mRNA was significantly up-regulated at one hour (h), 12 h and 24 h after stimulation with both V. anguillarum and M. lysodeikticu. The expression level of FcMyD88 protein was 2-fold up-regulated at 12 h post injection (hpi) of inactivated V. anguillarum while it didn't change after M. lysodeikticu injection during this period. After WSSV injection, the expression level of FcMyD88 mRNA remained relatively constant, while the FcMyD88 protein was significantly up-regulated at 12 and 24 hpi. These results suggested that the MyD88-dependent signaling pathway could be involved in the defense of both bacteria and WSSV infection.