Disruption of clathrin-dependent trafficking results in the failure of grass carp reovirus cellular entry.
Disruption of clathrin-dependent trafficking results in the failure of grass carp reovirus cellular entry.
复制标题
网格蛋白依赖性运输的破坏导致草鱼呼肠孤病毒无法进入细胞
DOI：
10.1186/s12985-016-0485-7
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发表时间：
2016-02-16
期刊：
影响因子：
4.8
通讯作者：
Lu L
中科院分区：
文献类型：
--
作者：
Wang H;Liu W;Yu F;Lu L
草鱼呼肠孤病毒（GCRV）是养殖草鱼（Ctenopharyngon idellus）病毒性出血性疾病的病原。GCRV是呼肠孤病毒科水膜病毒属的一种非包膜双链RNA病毒，编码7种结构蛋白（VP1-VP7）和5种非结构蛋白（NS80、NS38、NS31、NS26和NS16）。迄今为止，人们对GCRV进入CIK细胞（CIK）的机制知之甚少。结果我们研究了GCRV在CIK细胞中的内化机制。我们的研究结果表明氯丙嗪可以抑制GCRV感染，氯丙嗪是网格蛋白介导的内吞作用的特异性抑制剂。在感染早期，共聚焦显微镜观察到GCRV病毒粒子与内源性网格蛋白共定位。此外，GCRV对CIK细胞的感染依赖于内体的酸化。这表明，在使用溶酶促体药物氯喹或氯化铵进行预防性治疗后，病毒感染得到了显著的抑制。此外，动力蛋白活性的干扰阻断了GCRV的进入，这证实了网格蛋白介导的内吞作用依赖动力蛋白的性质。结论GCRV可能以ph依赖的方式通过网格蛋白介导的内吞作用进入CIK细胞。此外，动力蛋白对病毒有效进入至关重要。
BackgroundGrass carp reovirus (GCRV) is responsible for viral hemorrhagic disease in cultured grass carp (Ctenopharyngon idellus). GCRV is a non-enveloped, double-stranded RNA virus in the genusAquareovirus, of the familyReoviridae, which encodes seven structural proteins (VP1-VP7) and five nonstructural proteins (NS80, NS38, NS31, NS26, and NS16). To date, the mechanism of GCRV entry into CIKCtenopharyngon idelluskidney (CIK) cells remains poorly understood.ResultsHere, we present a study of the GCRV internalization mechanism in CIK cells. Our results indicated that GCRV infection was inhibited by chlorpromazine, the specific inhibitor for clathrin-mediated endocytosis. Colocalization of GCRV virions with endogenous clathrin was observed during early infection by confocal microscopy. Moreover, GCRV infection of CIK cells depended on acidification of the endosome. This was indicated by significant inhibition of viral infection following prophylactic treatment with the lysosomotropic drugs chloroquine or ammonium chloride. In addition, the disturbance of dynamin activity blocked GCRV entry, which confirmed the dynamin-dependent nature of clathrin-mediated endocytosis.ConclusionOur findings suggest that GCRV might enter CIK cells via clathrin-mediated endocytosis in a pH-dependent manner. Additionally, dynamin is critical for efficient viral entry.