Studies on betanodavirus non-structural protein B1 how to regulate the oxidative stresses in fish cell lines

• Main Authors: Heng-DaoLin, 林衡道
• Other Authors: Jiann-Ruey Hong
• Format: Others
• Language: zh-TW
• Published: 2013
• Online Access: http://ndltd.ncl.edu.tw/handle/36483187060182673047

碩士 === 國立成功大學 === 生物科技研究所碩博士班 === 101 === As we known, some fish species are infected by betanodavirus such as grouper, which results in severe mortality and significant economic losses on the aquaculture industry. In previous studies, RGNNV can induce necrotic cell death via mitochondrial membrane (MMP) loss in grouper cells. The RGNNV B1 is belonging to early expression gene which plays a crucial role on anti-apoptotic function and reduces viral replication. But, whether B1 overexpression can regulate ROS is still unknown. And how to reduce cell death with virus infection is also unclear. In this study, we found that the B1 can induce superoxide anion (O2- ) production at 12h. And then the B1 can up-regulate some anti-oxidant enzymes such as Cu/Zn SOD, Mn SOD, catalase and arrest the cell cycle via oxidative stress response. On the other hand, the B1 can reduce cell death with treat H2O2, tBHQ (H2O2 inducer) and Act D (Actinomycin D), an apoptosis inducer. Furthermore, we still don’t know whether B1 protein targeting into nucleus is correlative to anti-death function. In the result, we found that B1 targeting into nucleus can correlate to B1 function. Finally, we analyzed the transcription level by cDNA microarray after EYFP-B1 transfection and try to find some hint for ROS and anti-cell death studies. In the result, the B1 protein may play a multifunction roles, which not only arrest cell cycle but also induce ROS production and cell death. This first finding may provide an insight into the molecular pathogenesis of betanodavirus infection in future. Keywords: Betanodavirus, nonstructural protein B1, reactive oxygen species (ROS), oxidative stress respone, anti-oxidant enzyme, cell cycle arrest, anti-apoptosis.