Abstract：
The constitutive photomorphogenesis 9 (COP9) signalosome (CSN) typically composing of eight subunits (CSN1-8) mediates the process of deneddylation and deubiquitination. The fifth subunit of COP9 signalosome, CSN5, has special characteristics compared with the other seven subunits, and plays vital roles in the deneddylation activity and diverse cellular processes. However, the role of CSN5 in antiviral immunity is not clear. In this study, we identified 8 subunits (CSN1-8) of COP9 signalosome in shrimp Marsupenaeus japonicus. CSN1-6 were existed in all tested tissues, but CSN7-CSN8 were not detected in hepatopancreas. After WSSV challenged, the expression level of Csn1 to Csn4, and Csn6 to Csn8 were highly decreased, but the expression level of Csn5 was conspicuously increased in shrimp challenged by white spot syndrome virus (WSSV). The CSN5 was recombinantly expressed in Escherichia coli and its polyclonal antibody was prepared. The expression level of CSN5 was conspicuously increased at RNA and protein levels in the shrimp challenged by WSSV. After knockdown of Csn5 by RNA interference, the WSSV replication was obviously increased in shrimp. When injected the recombinant protein of CSN5 with the membrane penetrating peptide into shrimp, WSSV replication was inhibited and the survival rate of shrimp was significantly improved compared with control. We further analyzed the expression of antimicrobial peptides (AMPs) in Csn5-RNAi shrimp, and the results showed that the expression of several AMPs was declined significantly. These results indicate that CSN5 inhibits replication of WSSV via regulating expression of AMPs in shrimp, and the recombinant CSN5 might be used in shrimp aquaculture for the white spot syndrome disease control.
摘要：
通常由八个亚基（CSN1-8）组成的组成型光形态发生9（COP9）信号体（CSN）介导去修饰和去泛素化的过程。COP9信号体的第五亚基，CSN5，与其他七个亚基相比具有特殊的特点，并在去乙酰化活性和多种细胞过程中起着至关重要的作用。然而,CSN5在抗病毒免疫中的作用尚不清楚。在这项研究中,我们在日本对虾中鉴定了COP9信号体的8个亚基（CSN1-8）。CSN1-6存在于所有测试组织中，但在肝胰腺中未检测到CSN7-CSN8。在WSSV挑战之后,Csn1对Csn4和Csn6对Csn8的表达水平高度降低，但是Csn5的表达水平在受到白斑综合症病毒（WSSV）攻击的虾中明显增加。在大肠杆菌中重组表达CSN5并制备其多克隆抗体。在受到WSSV攻击的虾中，CSN5的表达水平在RNA和蛋白质水平上明显增加。通过RNA干扰敲低Csn5后，对虾的WSSV复制明显增加。当将CSN5的重组蛋白与膜穿透肽一起注射到虾中时，与对照相比，WSSV复制受到抑制，虾的存活率明显提高。我们进一步分析了抗菌肽（AMPs）在Csn5-RNAi虾中的表达，结果表明，几种AMPs的表达明显下降。这些结果表明，CSN5通过调节虾中AMPs的表达来抑制WSSV的复制，重组CSN5可能用于对虾养殖，以控制白斑综合征。
