Soponpong Suthinee, Amparyup Piti, Tassanakajon Anchalee
Center of Excellence for Molecular Biology and Genomics of Shrimp, Department of Biochemistry, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand.
National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Pathumthani 12120, Thailand.
Dev Comp Immunol. 2018 Apr;81:291-302. doi: 10.1016/j.dci.2017.12.013. Epub 2017 Dec 14.
DEAD (Asp-Glu-Ala-Asp)-box polypeptide 41 (DDX41), a receptor belonging to the DExD family, has recently been identified as an intracellular DNA sensor in vertebrates. Here, we report on the identification and functional characterization of PmDDX41, the first cytosolic DNA sensor in shrimp. By searching a Penaeus monodon expressed sequence tag (EST) database (http://pmonodon.biotec.or.th), three cDNA fragments exhibiting similarity to DDX41 in various species were identified and assembled, resulting in a complete open reading frame of PmDDX41 that contains 1863-bp and encodes a putative protein of 620 amino acids. PmDDX41 shares 83% and 79% similarity to DDX41 homolog from the bee Apis florea and fruit fly Drosophila melanogaster, respectively and contains three conserved domains in the protein: DEADc domain, HELICc domain, and zinc finger domain. The transcript of PmDDX41 was detected in all tested tissues and was up-regulated upon infection with a DNA virus, white spot syndrome virus (WSSV). However, PmDDX41 mRNA expression was not significantly changed and down-regulated in response to a bacterium, Vibrio harveyi, or an RNA virus, yellow head virus (YHV), respectively, compared with the control phosphate-buffered saline-injected shrimp. Furthermore, the suppression of PmDDX41 by dsRNA-mediated gene silencing resulted in more rapid death of WSSV-infected shrimp and a significant decrease in the mRNA expression levels of several immune-related genes (PmIKKβ, PmIKKɛ, PmRelish, PmCactus, PmDorsal, PmPEN3, PmPEN5, and ALFPm6). These results suggest that PmDDX41 is involved in the antiviral response, probably via a DNA-sensing pathway that is triggered through the IκB kinase complex and leads to the activation of several immune-related genes.
DEAD(天冬氨酸-谷氨酸-丙氨酸-天冬氨酸)盒多肽41(DDX41)是一种属于DExD家族的受体,最近被鉴定为脊椎动物中的一种细胞内DNA传感器。在此,我们报告了对虾中首个胞质DNA传感器PmDDX41的鉴定及其功能特性。通过搜索斑节对虾表达序列标签(EST)数据库(http://pmonodon.biotec.or.th),鉴定并组装了三个与不同物种中的DDX41具有相似性的cDNA片段,从而得到了PmDDX41的一个完整开放阅读框,其长度为1863 bp,编码一个推定的由620个氨基酸组成的蛋白质。PmDDX41与来自蜜蜂小蜜蜂和果蝇黑腹果蝇的DDX41同源物分别具有83%和79%的相似性,并且在该蛋白质中包含三个保守结构域:DEADc结构域、HELICc结构域和锌指结构域。在所有测试组织中均检测到了PmDDX41的转录本,并且在用DNA病毒白斑综合征病毒(WSSV)感染后其表达上调。然而,与注射对照磷酸盐缓冲盐水的虾相比,在分别受到细菌哈维氏弧菌或RNA病毒黄头病毒(YHV)刺激时,PmDDX41的mRNA表达没有显著变化,反而下调。此外,通过双链RNA介导的基因沉默抑制PmDDX41会导致WSSV感染的虾更快死亡,并且几种免疫相关基因(PmIKKβ、PmIKKɛ、PmRelish、PmCactus、PmDorsal、PmPEN3、PmPEN5和ALFPm6)的mRNA表达水平显著降低。这些结果表明,PmDDX41可能通过由IκB激酶复合物触发并导致几种免疫相关基因激活的DNA传感途径参与抗病毒反应。