Newly identified PcToll4 regulates antimicrobial peptide expression in intestine of red swamp crayfish Procambarus clarkii
Newly identified PcToll4 regulates antimicrobial peptide expression in intestine of red swamp crayfish Procambarus clarkii
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新发现的 PcToll4 调节红沼泽克氏原螯虾肠道中抗菌肽的表达
DOI：
10.1016/j.gene.2017.02.018
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发表时间：
2017
期刊：
影响因子：
3.5
通讯作者：
Qian Ren
中科院分区：
文献类型：
--
作者：
Ying Huang;Li Tingting;Min Jin;Yin Shaowu;Hui Kai Min;Qian Ren
Toll或Toll样受体(TLRs)在启动针对病原体的先天免疫反应中具有重要作用。本研究首次从克氏原虾肠道转录组中鉴定出一个新的Toll基因PcToll4。PcToll4全长4849hp，开放阅读框3036hp，编码1011个氨基酸。PcTol14含有一个信号肽、13个LRR结构域、3个LRR TYP结构域、2个LRR CT结构域、1个LRR NT结构域、1个跨膜区和1个TIR结构域。定量RT-PCR分析表明，PcTol14在所有受试组织中均有表达，白斑综合征病毒(WSSV)攻击后，PcTol14在肠道中的表达显著上调。PcToll4在果蝇Schneider 2(S2)细胞中的过表达可激活果蝇体内的抗菌肽(AMPs)，包括麦氏杆菌素(Metchnikowin)、屈曲霉素A、阿塔卡菌素A(AttacinA)和对虾抗菌素-4(HRMPPenaeidin-4)。SiRNA干扰结果还表明，PcTol14对小龙虾肠道中5种抗脂多糖因子(ALF)的表达具有调节作用。我们的发现表明，PcTol14基因对克氏肺孢子虫的先天免疫反应很重要，因为该基因调节抗WSSV的AMP的表达。
Tolls or Toll-like receptors (TLRs) have an essential role in initiating innate immune responses against pathogens. In this study, a novel Toll gene,PcToll4, was first identified from the intestinal transcriptome of the freshwater crayfish, Procambarus clarkii. ThePcToll4cDNA is 4849 bp long with a 3036 bp open reading frame that encodes a 1011-amino acid protein. PcToll4 contains a signal peptide, 13 LRR domains, 3 LRR TYP domains, 2 LRR CT domains, an LRR NT domain, a transmembrane region, and a TIR domain. Quantitative RT-PCR analysis revealed thatPcToll4mRNA was detected in all tested tissues, and the expression ofPcToll4in the intestine was significantly upregulated after white spot syndrome virus (WSSV) challenge. Overexpression of PcToll4 inDrosophilaSchneider 2 (S2) cells activates the antimicrobial peptides (AMPs) ofDrosophila, includingmetchnikowin,drosomycin,attacinA, and shrimpPenaeidin-4. Results of RNA interference by siRNA also showed thatPcToll4regulates the expressions of 5 anti-lipopolysaccharide factors (ALFs) in the intestine of crayfish. Our findings suggest thatPcToll4is important for the innate immune responses ofP.clarkiibecause this gene regulates the expressions of AMPs against WSSV.