White spot syndrome virus enters crayfish hematopoietic tissue cells via clathrin-mediated endocytosis
White spot syndrome virus enters crayfish hematopoietic tissue cells via clathrin-mediated endocytosis
复制标题
DOI：
10.1016/j.virol.2015.08.034
复制
发表时间：
2015-12-01
期刊：
影响因子：
3.7
通讯作者：
Yang, Feng
中科院分区：
文献类型：
--
作者：
Huang, Jiajun;Li, Fang;Yang, Feng
白斑综合征病毒（WSSV）是养殖对虾的主要病原体。然而，其进入的机制仍然知之甚少。本研究通过分析小龙虾血细胞组织（HPT）细胞对WSSV的内化，我们发现WSSV病毒粒子被具有网格蛋白包被凹坑特征的细胞膜吞噬，然后内化到包被的细胞质囊泡中。进一步研究表明，氯丙嗪（CPZ）对WSSV内化有显著抑制作用，而染料木素对WSSV内化无显著抑制作用。内化病毒粒子与内源性网格蛋白和转铁蛋白共定位，转铁蛋白依赖于网格蛋白的摄取。防止内体被氯化铵（NH4Cl）或氯喹（CQ）酸化，也显著减少了WSSV的进入。此外，动力蛋白活性的紊乱或膜胆固醇的消耗也会阻碍WSSV的摄取。这些数据表明，WSSV通过网格蛋白介导的内吞作用以ph依赖的方式进入小龙虾HPT细胞，膜胆固醇和动力蛋白对病毒有效进入至关重要。(C) 2015爱思唯尔公司版权所有。
White spot syndrome virus (WSSV) is a major pathogen of aquacultured shrimp. However, the mechanism of its entry remains poorly understood. In this study, by analyzing the internalization of WSSV using crayfish hematopoietic tissue (HPT) cells, we showed that WSSV virions were engulfed by cell membrane invaginations sharing the features of clathrin-coated pits and then internalized into coated cytoplasmic vesicles. Further investigation indicated that WSSV internalization was significantly inhibited by chlorpromazine (CPZ) but not genistein. The internalized virions were colocalized with endogenous clathrin as well as transferrin which undergoes clathrin-dependent uptake. Preventing endosome acidification by ammonium chloride (NH4Cl) or chloroquine (CQ) dramatically reduced WSSV entry as well. Moreover, disturbance of dynamin activity or depletion of membrane cholesterol also blocked WSSV uptake. These data indicate that WSSV enters crayfish HPT cells via clathrin-mediated endocytosis in a pH-dependent manner, and membrane cholesterol as well as dynamin is critical for efficient viral entry. (C) 2015 Elsevier Inc. All rights reserved.