Yang Feifei, Li Shihao, Li Fuhua, Xiang Jianhai
Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China.
Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China; Laboratory for Marine Biology and Biotechnology, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266237, China.
Dev Comp Immunol. 2018 Apr;81:303-311. doi: 10.1016/j.dci.2017.12.018. Epub 2017 Dec 21.
White spot syndrome virus (WSSV) is a major viral pathogen in global shrimp farming, causing huge economic damage. Through penetrating the outer surface of the target tissues, WSSV enters into the cells of the target tissue to complete the replication process in the host. In the present study, a cuticle protein gene from Litopenaeus vannamei, designated as LvAMP13.4, was identified and proved to be involved in WSSV invasion. The deduced amino acid sequence of LvAMP13.4 contained a signal peptide and a conserved chitin-binding domain type 4 (ChBD4). This cuticle protein gene was mainly expressed in stomach, gill and epidermis. The expression level of LvAMP13.4 was significantly changed during WSSV infection. Silencing of LvAMP13.4 by dsRNA interference apparently reduced the mortality rate and the WSSV copy number in shrimp upon WSSV infection. Furthermore, yeast two-hybrid system and Co-IP assay were performed to confirm that LvAMP13.4 could interact with the major envelop protein VP24 of WSSV. These data indicated that LvAMP13.4 was involved in the invasion process of WSSV through interaction with VP24. The present results could provide new insights for us in understanding the role of host cuticle proteins during virus invasion.
白斑综合征病毒（WSSV）是全球对虾养殖中的一种主要病毒病原体，会造成巨大的经济损失。WSSV通过穿透靶组织的外表面进入靶组织细胞，在宿主体内完成复制过程。在本研究中，从凡纳滨对虾中鉴定出一种表皮蛋白基因，命名为LvAMP13.4，并证明其参与WSSV的入侵。LvAMP13.4推导的氨基酸序列包含一个信号肽和一个保守的4型几丁质结合结构域（ChBD4）。该表皮蛋白基因主要在胃、鳃和表皮中表达。在WSSV感染期间，LvAMP13.4的表达水平发生显著变化。通过dsRNA干扰沉默LvAMP13.4，明显降低了WSSV感染后对虾的死亡率和WSSV拷贝数。此外，通过酵母双杂交系统和免疫共沉淀试验证实，LvAMP13.4可与WSSV的主要包膜蛋白VP24相互作用。这些数据表明，LvAMP13.4通过与VP24相互作用参与WSSV的入侵过程。本研究结果可为我们理解宿主表皮蛋白在病毒入侵过程中的作用提供新的见解。
