石斑鱼虹彩病毒（GIV-R）关联宿主蛋白GRP78在病毒复制中的功能研究

项目名称： 石斑鱼虹彩病毒（GIV-R）关联宿主蛋白GRP78在病毒复制中的功能研究

项目编号： No.31502211

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 农业科学

项目作者： 马红玲

作者单位： 中国水产科学研究院南海水产研究所

项目金额： 21万元

中文摘要： 葡萄糖调节蛋白GRP78是内质网的标志性分子伴侣，在内质网应激（ERS）发生时发挥重要的调节功能。在病毒感染触发的ERS反应中，多数研究显示宿主GRP78可被病毒挟持参与感染性病毒颗粒的形成，从而有利于病毒的复制进程。蛙病毒属石斑鱼虹彩病毒（GIV-R）是养殖石斑鱼的重要病原，我们的研究已经证实GRP78作为宿主来源病毒囊膜蛋白参与了成熟GIV-R粒子的形成，但其在GIV-R复制中的角色还不清楚。本研究拟通过分析GRP78等ERS的核心关联蛋白在GIV-R感染宿主中的表达时序，研究GIV-R感染与宿主ERS发生的联系；构建过表达或沉默GRP78研究体系，评估GRP78对GIV-R复制的影响；酵母双杂交实验发掘与GPR78相互作用的病毒蛋白。研究结果将揭示GRP78在GIV-R复制中的角色，对于深入理解GIV-R的致病机制具有重要科学意义。

中文关键词： 病毒病；感染；复制；致病因子；结构

英文摘要： Glucose regulated protein 78 (GRP78), one of the landmark molecule chaperones in endoplasmic reticulum (ER), plays an important regulation role when ER stress happens. In study of the interaction relationship between high animal/human viruses and their host proteins, the majority of studies showed that GRP78 was always hijacked by virus to be assembled into maturation of productive infection virions and play a crucial role in viral infection or replication. In aquatic viruses, however, few studies have been carried out in GPR78 function. Grouper iridovirus belonging to genus Ranavirus (GIV-R) is an important causative agent in grouper industry in SE Asia including mainland China, Chinese Taiwan and Singapore etc. In our recent proteomic study of a newly isolated GIV-R from diseased cultured grouper juveniles in Hainan Island, GRP78 was confirmed for the first time as a host derived viral envelope in purified GIV-R virions. However, the functional role of GRP78 during GIV-R replication remains unclear. To explore the functional role of GRP78, in this proposal, quantitatively temporal expression of GRP78 and other ER associated core proteins in GIV-R-infected grouper cell line (GB11) and groupers will be determined to study the inter relationship between GIV-R infection and ERS occurrence; Research system of overexpression and knockdown of GRP78 will be established to assess the function of GRP78 in GIV-R replication; finally, yeast-two hybrid experiment will be performed to explore the possible interaction proteins between viral structural proteins and GRP78. The expected results will reveal the functional role of GRP78 in GIV-R replication, which will be of scientific importance for better understanding of the pathogenic mechanism of GIV-R.

英文关键词： Viral diseases;Infection;Replication;Pathogenic factor;Structure