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对小黄鱼小RNA转录组的比较分析揭示了微小RNA介导的Toll样受体信号通路对鳗弧菌感染反应的调控。

Comparative analysis of the small RNA transcriptomes of miiuy croaker revealed microRNA-mediated regulation of TLR signaling pathway response to Vibrio anguillarum infection.

作者信息

Xu Guoliang, Han Jingjing, Xu Tianjun

机构信息

Laboratory of Fish Biogenetics & Immune Evolution, College of Marine Science, Zhejiang Ocean University, Zhoushan, 316022, China.

Laboratory of Fish Biogenetics & Immune Evolution, College of Marine Science, Zhejiang Ocean University, Zhoushan, 316022, China.

出版信息

Fish Shellfish Immunol. 2016 May;52:248-57. doi: 10.1016/j.fsi.2016.03.011. Epub 2016 Mar 12.

Abstract

MicroRNAs (miRNAs) are an abundant class of endogenous noncoding small RNAs (sRNAs) that are partially complementary to their target messenger RNA (mRNA), which post-transcriptionally regulate various biological processes by repressing mRNA translation or inducing mRNA degradation. MiRNAs have been demonstrated to play crucial roles in the host response to infection by diverse pathogens. As an important bacterial pathogen, Vibrio anguillarum has been caused great economic losses in miiuy croaker aquaculture. To identify immune-related miRNAs of miiuy croaker in response to V. anguillarum infection, we constructed two sRNA libraries with or without bacterial infection. High-throughput deep sequencing and subsequent bioinformatic analysis identified 241 conserved and 137 novel miRNA precursors in miiuy croaker based on its whole genome sequences, encoding 293 and 124 mature miRNAs, respectively. Then we compared the expression patterns of miRNAs in the two libraries. There were significant differences in the expression of 12 miRNAs between the infection group (IG) and control group (CG). Further, the expressions of six miRNAs were validated by real-time quantitative PCR. The target gene prediction and function analysis were conducted for the 12 differential miRNAs. This analysis revealed that these miRNAs participated in the regulation multiple immune-related signaling pathways. Transcription factors in TLR signaling, such as AP-1, IRF5, NF-κB and IRF3, were activated by these miRNAs via post-transcriptionally regulating the expression of TLRs and TLR-associated signaling proteins, inducing effective host immune response to eradicate infectious pathogens. This is the first study of the identification and characterization of miiuy croaker miRNAs in response to V. anguillarum infection. The comprehensive analysis of the expression of miRNAs and the target gene and function prediction of differently expressed miRNAs may help to understand the regulatory mechanisms of miRNA in fish during the interaction between host and bacterial pathogens.

摘要

微小RNA(miRNA)是一类丰富的内源性非编码小RNA(sRNA),它们与其靶信使RNA(mRNA)部分互补,通过抑制mRNA翻译或诱导mRNA降解在转录后调节各种生物学过程。已证明miRNA在宿主对多种病原体感染的反应中起关键作用。鳗弧菌作为一种重要的细菌病原体,给大黄鱼养殖带来了巨大的经济损失。为了鉴定大黄鱼对鳗弧菌感染的免疫相关miRNA,我们构建了有无细菌感染的两个sRNA文库。基于大黄鱼全基因组序列,通过高通量深度测序和后续生物信息学分析,鉴定出241个保守的和137个新的miRNA前体,分别编码293个和124个成熟miRNA。然后我们比较了两个文库中miRNA的表达模式。感染组(IG)和对照组(CG)之间有12个miRNA的表达存在显著差异。此外,通过实时定量PCR验证了6个miRNA的表达。对这12个差异miRNA进行了靶基因预测和功能分析。该分析表明这些miRNA参与了多种免疫相关信号通路的调节。这些miRNA通过转录后调节TLR和TLR相关信号蛋白的表达,激活TLR信号通路中的转录因子,如AP-1、IRF5、NF-κB和IRF3,诱导宿主产生有效的免疫反应以根除感染性病原体。这是首次对大黄鱼响应鳗弧菌感染的miRNA进行鉴定和表征的研究。对miRNA表达的综合分析以及差异表达miRNA的靶基因和功能预测可能有助于理解鱼类在宿主与细菌病原体相互作用过程中miRNA的调控机制。

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