Characterization of Argonaute2 gene from black tiger shrimp (Penaeus monodon) and its responses to immune challenges.
Characterization of Argonaute2 gene from black tiger shrimp (Penaeus monodon) and its responses to immune challenges.
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DOI：
10.1016/j.fsi.2013.11.010
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发表时间：
2014
影响因子：
4.7
通讯作者：
Li-Shi Yang;Xiaolan Li;Song Jiang;L. Qiu;F. Zhou;Wenjing Liu;Shi-gui Jiang
中科院分区：
文献类型：
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作者：
Li-Shi Yang;Xiaolan Li;Song Jiang;L. Qiu;F. Zhou;Wenjing Liu;Shi-gui Jiang
Argonaute2与短向导RNA（microRNA或短干扰RNA）结合，向导RNA将RISC引导至互补mRNA，这些互补mRNA是RISC介导的基因沉默的靶点。本文对斑节对虾Argonaeus monodon中Argonaute 2基因（命名为PmAgo 2）进行了鉴定和分析。PmAgo2基因的5′端非翻译区（UTR）为106 bp，开放阅读框（ORF）为2616 bp，3′端非翻译区（UTR）为123 bp。预测的PmAgo2蛋白为99.4 KDa，理论等电点为9.54。PmAgo 2与日本囊对虾Argonaeus 2和凡纳滨对虾Argonaeus 2的氨基酸序列相似性最高，分别为69.0%和68.5%。系统发育分析表明，PmAgo2与虾Argonaute2聚为一类，与昆虫类关系较近。实时荧光定量PCR结果显示，PmAgo2在除眼柄外的所有组织中均有表达，在淋巴液和血细胞中表达量较高。金黄色葡萄球菌（Staphylococcus aureus）和白色斑点综合征病毒（White Spot Syndrome Virus，WSSV）能显著上调肝胰腺中PmAgo 2的mRNA表达。此外，我们的研究还证实了dsRNA和ssRNA同源poly（I：C）和R848激活PmAgo2的表达。结果表明，PmAgo2对细菌感染和病毒感染均有应答，尤其是与其他siRNA途径的核心成员一起，在黑虎虾体内诱导了ssRNA介导的RNAi。
Argonaute2 binds to a short guide RNA (microRNA or short interfering RNA) and guides RNAs direct RISC to complementary mRNAs that are targets for RISC-mediated gene silencing. Here we identified and characterized Argonaute2 from black tiger shrimpPenaeus monodon(designated as PmAgo2). The full-length cDNA of PmAgo2 contained a 5′ untranslated region (UTR) of 106 bp, an open reading frame (ORF) of 2616 bp and a 3′ UTR of 123 bp. The predicted PmAgo2 protein is 99.4 KDa with the theoretical isoelectric point of 9.54. PmAgo2 shared the highest similarity of amino acid withMarsupenaeus japonicusArgonaute2 andLitopenaeus vannameiArgonaute2, at 69.0% and 68.5%, respectively. Phylogenic analysis showed PmAgo2 clustered with shrimp Argonaute2, and closed to the group of insects. Real-time quantitative PCR showed that PmAgo2 was widely expressed in almost all examined tissues except eyestalk, with high expression in lymph and haemocyte. mRNA expression also revealed that PmAgo2 was significantly up-regulated byStaphylococcus aureusand White Spot Syndrome Virus (WSSV) in hepatopancreas. Furthermore, our study also confirmed that dsRNA and ssRNA homologous poly (I:C) and R848 activated the expression of PmAgo2. The result indicated that PmAgo2 responded to both bacterial infection and viral infection, especially, it may induce an ssRNA-mediated RNAi with other core members of siRNA pathway in black tiger shrimp.