Pang Zhenguo, Kim Su-Kyoung, Yu Jiaping, Jang In-Kwon
National Fisheries Research & Development Institute, Ulwang-dong 707, Jung-gu, Incheon 400-420, South Korea.
Fish Shellfish Immunol. 2014 Aug;39(2):158-67. doi: 10.1016/j.fsi.2014.04.026. Epub 2014 May 9.
Prophenoloxidase activating enzyme 2 (PPAE2), which belongs to the second PPAE family of prawns, was isolated from white shrimp Litopenaeus vannamei. The currently identified lvPPAE2 and lvPPAE1 from our former report were taken as model candidates to analyze the relationship of the two shrimp PPAE families as well as the regulation mechanism of shrimp PPAEs. The tissue expression of lvPPAE2 was more ubiquitous than lvPPAE1. The mRNA abundance of lvPPAE2 was about 10 percent of lvPPAE1 in co-existed tissues. When challenged with Vibrio harveyi. LvPPAE2 showed a distinct transcriptional regulation pattern compared to lvPPAE1. Silence of lvPPAE2 significantly increased shrimp's susceptibility to V. harveyi, suggesting the lvPPAE2 plays essential role in shrimp host defense. A novel PPAE specific compensatory over expression feature was found in the two lvPPAEs. Single gene specific silence of lvPPAE1 and lvPPAE2 resulted in a significant difference in reduction of hemolymph PO activity. Double silence of the two lvPPAEs failed to cause a further reduction on PO activity or shrimp mortality to bacteria, despite that double silence sufficiently suppressed both of the two lvPPAEs. Our findings suggest both lvPPAEs contribute to shrimp melanization cascade and host defense against bacteria. Distinct regulation pattern corresponding to the same pathogen invasion suggests the two lvPPAEs are actually under different regulation ways. A novel PPAE specific compensatory over expression mechanism found in our study offered us a clue in understanding the robustness of shrimp innate immunity and network of crustacean proPO activating system.
酚氧化酶原激活酶2(PPAE2)属于对虾的第二个PPAE家族,是从凡纳滨对虾中分离得到的。将我们之前报告中目前已鉴定的lvPPAE2和lvPPAE1作为模型候选物,以分析对虾两个PPAE家族之间的关系以及对虾PPAE的调控机制。lvPPAE2的组织表达比lvPPAE1更广泛。在共存组织中,lvPPAE2的mRNA丰度约为lvPPAE1的10%。当用哈维氏弧菌攻击时,与lvPPAE1相比,LvPPAE2表现出明显不同的转录调控模式。lvPPAE2基因沉默显著增加了对虾对哈维氏弧菌的易感性,这表明lvPPAE2在对虾宿主防御中起重要作用。在这两种lvPPAE中发现了一种新的PPAE特异性补偿性过表达特征。lvPPAE1和lvPPAE2的单基因特异性沉默导致血淋巴酚氧化酶活性降低存在显著差异。尽管两种lvPPAE的双基因沉默充分抑制了它们,但这两种lvPPAE的双基因沉默未能进一步降低酚氧化酶活性或对虾对细菌的死亡率。我们的研究结果表明,两种lvPPAE都有助于对虾的黑化级联反应和宿主对细菌的防御。对应于相同病原体入侵的不同调控模式表明,这两种lvPPAE实际上处于不同的调控方式。我们研究中发现的一种新的PPAE特异性补偿性过表达机制为我们理解对虾先天免疫的稳健性和甲壳动物酚氧化酶原激活系统的网络提供了线索。