Key Laboratory of Exploration and Utilization of Aquatic Resources, Ministry of Education, National Demonstration Center for Experimental Fisheries Science Education, College of Fisheries and Life Science, Shanghai Ocean University, Shanghai, 201306, China; Key Laboratory of South China Sea Fishery Resources Exploitation and Utilization, Ministry of Agriculture and Rural Affairs, South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, 510300, Guangzhou, Guangdong Province, China.
Key Laboratory of South China Sea Fishery Resources Exploitation and Utilization, Ministry of Agriculture and Rural Affairs, South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, 510300, Guangzhou, Guangdong Province, China; Sanya Tropical Fisheries Research Institute, Sanya, Hainan Province, China.
Mol Immunol. 2021 May;133:77-85. doi: 10.1016/j.molimm.2021.02.002. Epub 2021 Feb 23.
Tripartite motif-containing 9 (TRIM9) has been demonstrated to exert important roles in regulation of innate immune signaling. In this study, a novel TRIM9 homolog was identified from Penaeus monodon (named PmTRIM9). The open reading frame (ORF) of PmTRIM9 was 2064 bp, which encoding a 687-amino-acid polypeptide. Following Vibrio parahaemolyticus challenge, the expression levels of PmTRIM9 mRNA were significantly down-regulated in tested tissues. RNA interference and recombinant protein injection experiments were performed to explore the function of PmTRIM9, and the results showed it could facilitate V. parahaemolyticus replication and lead P. monodon more vulnerable to V. parahaemolyticus challenge. The dual-luciferase reporter assay showed that PmTRIM9 possessed the ability to inhibit the promoter activity in HEK293 T cells. Silencing of PmTRIM9 could increase the expression of the major NF-κB transcription factor, PmRelish. Further studies showed that knockdown of PmRelish promoted the V. parahaemolyticus infection and decreased the expression of specific antimicrobial peptides (AMPs), including PmCRU5, PmCRU7, PmALF6, PmALF3, PmLYZ and PmPEN5. However, knockdown of PmTRIM9 increased expression levels of the same AMPs, but except for PmCRU5, indicating that PmTRIM9 may negatively regulate the PmRelish-mediated expression of AMPs. All these results suggest that PmTRIM9 was involved in facilitating V. parahaemolyticus infection by inhibition of Relish pathway in P. monodon.
三结构域蛋白 9（TRIM9）已被证明在调节先天免疫信号中发挥重要作用。在本研究中，从斑节对虾（命名为 PmTRIM9）中鉴定出一种新型 TRIM9 同源物。PmTRIM9 的开放阅读框（ORF）为 2064 bp，编码 687 个氨基酸的多肽。在副溶血弧菌攻毒后，检测组织中 PmTRIM9 mRNA 的表达水平显著下调。进行了 RNA 干扰和重组蛋白注射实验，以探索 PmTRIM9 的功能，结果表明它可以促进副溶血弧菌的复制，使斑节对虾更容易受到副溶血弧菌的攻击。双荧光素酶报告基因实验显示 PmTRIM9 具有抑制 HEK293T 细胞启动子活性的能力。沉默 PmTRIM9 可以增加主要 NF-κB 转录因子 PmRelish 的表达。进一步的研究表明，PmRelish 的敲低促进了副溶血弧菌的感染，并降低了特定抗菌肽（AMPs）的表达，包括 PmCRU5、PmCRU7、PmALF6、PmALF3、PmLYZ 和 PmPEN5。然而，PmTRIM9 的敲低增加了相同 AMPs 的表达水平，但除了 PmCRU5 外，这表明 PmTRIM9 可能通过负调控 Relish 途径来负调控 PmRelish 介导的 AMPs 表达。所有这些结果表明，PmTRIM9 通过抑制斑节对虾中的 Relish 途径参与促进副溶血弧菌感染。
