Huang Mingzhu, Liu Yuan, Xie Chenying, Wang Wei-Na
Key Laboratory of Ecology and Environmental Science in Guangdong Higher Education, Guangdong Provincial Key Laboratory for Healthy and Safe Aquaculture, College of Life Science, South China Normal University, Guangzhou 510631, PR China.
Key Laboratory of Ecology and Environmental Science in Guangdong Higher Education, Guangdong Provincial Key Laboratory for Healthy and Safe Aquaculture, College of Life Science, South China Normal University, Guangzhou 510631, PR China.
Fish Shellfish Immunol. 2015 May;44(1):180-6. doi: 10.1016/j.fsi.2015.02.022. Epub 2015 Feb 20.
DJ-1 was first identified as an oncogene that transformed mouse NIH3T3 cells in cooperation with activated Ras. It has since exhibited a variety of functions in a range of organisms. In this study, the DJ-1 gene in Litopenaeus vannamei (LvDJ-1) was identified and characterized. A recombinant protein LvDJ-1 was produced in Pichia pastoris. LvDJ-1 expression in vivo was knocked down by dsRNA-mediated RNA interference (RNAi), which led to significantly decreased levels of LvDJ-1 mRNA and protein. When the L. vannamei were challenged with RNAi and Vibrio alginolyticus, the transcription and expression of copper zinc superoxide dismutase (LvCZSOD) in the hepatopancreas were dramatically lower in shrimp with knocked down LvDJ-1 than in controls. Transcription and expression of P53 (LvP53) were significantly higher in shrimp lacking LvDJ-1 than in controls. Hepatopancreas samples were analyzed using real time polymerase chain reaction and Western blot. Moreover, blood samples from the shrimp, assessed with flow cytometry, showed significant increases in respiratory burst and apoptosis in those lacking LvDJ-1 compared to the controls. Cumulative mortality in the shrimp lacking LvDJ-1 was significantly different from that in the control group after challenge with V. alginolyticus. Altogether, the results prove that LvDJ-1 regulates apoptosis and antioxidant activity, and that these functions play an important role in L. vannamei resistance against V. alginolyticus.
DJ-1最初被鉴定为一种癌基因,它与激活的Ras协同作用可转化小鼠NIH3T3细胞。此后,它在一系列生物体中表现出多种功能。在本研究中,对凡纳滨对虾(LvDJ-1)中的DJ-1基因进行了鉴定和表征。在毕赤酵母中产生了重组蛋白LvDJ-1。通过dsRNA介导的RNA干扰(RNAi)敲低了体内的LvDJ-1表达,这导致LvDJ-1 mRNA和蛋白水平显著降低。当用RNAi和溶藻弧菌对凡纳滨对虾进行攻毒时,LvDJ-1被敲低的对虾肝胰腺中铜锌超氧化物歧化酶(LvCZSOD)的转录和表达显著低于对照组。缺乏LvDJ-1的对虾中P53(LvP53)的转录和表达显著高于对照组。使用实时聚合酶链反应和蛋白质免疫印迹法对肝胰腺样本进行了分析。此外,用流式细胞术评估对虾的血样,结果显示,与对照组相比,缺乏LvDJ-1的对虾呼吸爆发和细胞凋亡显著增加。在用溶藻弧菌攻毒后,缺乏LvDJ-1的对虾的累积死亡率与对照组有显著差异。总之,结果证明LvDJ-1调节细胞凋亡和抗氧化活性,并且这些功能在凡纳滨对虾抵抗溶藻弧菌中起重要作用。