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电压依赖性阴离子通道在日本脑炎病毒感染昆虫细胞过程中发生重新分布。

Voltage dependent anion channel is redistributed during Japanese encephalitis virus infection of insect cells.

作者信息

Fongsaran Chanida, Phaonakrop Narumon, Roytrakul Sittiruk, Thepparit Chutima, Kuadkitkan Atichat, Smith Duncan R

机构信息

Institute of Molecular Biosciences, Mahidol University, Salaya Campus, 25/25 Phuttamonthol Sai 4 Road, Salaya, Nakhon Pathom 73170, Thailand.

Proteomics Research Laboratory, Genome Institute, National Science and Technology Development Agency, 113 Thailand Science Park, Phahonyothin Road, Khlong Nueng, Khlong Luang, Pathum Thani 12120, Thailand.

出版信息

ScientificWorldJournal. 2014;2014:976015. doi: 10.1155/2014/976015. Epub 2014 Jul 10.

Abstract

Despite the availability of an effective vaccine, Japanese encephalitis remains a significant cause of morbidity and mortality in many parts of Asia. Japanese encephalitis is caused by the Japanese encephalitis virus (JEV), a mosquito transmitted flavivirus. Many of the details of the virus replication cycle in mosquito cells remain unknown. This study sought to determine whether GRP78, a well-characterized flavivirus E protein interacting protein, interacted with JEV E protein in insect cells, and whether this interaction was mediated at the cell surface. GRP78 was shown to interact with JEV E protein by coimmunoprecipitation, and was additionally shown to interact with voltage dependent anion protein (VDAC) through the same methodology. Antibody inhibition experiments showed that neither GRP78 nor VDAC played a role in JEV internalization to insect cells. Interestingly, VDAC was shown to be significantly relocalized in response to JEV infection, and significant levels of colocalization between VDAC and GRP78 and VDAC and ribosomal L28 protein were seen in JEV infected but not uninfected cells. This is the first report of relocalization of VDAC in response to JEV infection and suggests that this may be a part of the JEV replication strategy in insect cells.

摘要

尽管有有效的疫苗,但日本脑炎在亚洲许多地区仍然是发病和死亡的重要原因。日本脑炎由日本脑炎病毒(JEV)引起,这是一种通过蚊子传播的黄病毒。病毒在蚊子细胞中的复制周期的许多细节仍然未知。本研究旨在确定GRP78(一种特征明确的黄病毒E蛋白相互作用蛋白)是否在昆虫细胞中与JEV E蛋白相互作用,以及这种相互作用是否在细胞表面介导。通过共免疫沉淀显示GRP78与JEV E蛋白相互作用,并通过相同方法额外显示其与电压依赖性阴离子蛋白(VDAC)相互作用。抗体抑制实验表明,GRP78和VDAC在JEV内化进入昆虫细胞过程中均不起作用。有趣的是,显示VDAC在JEV感染后发生显著的重新定位,并且在JEV感染而非未感染的细胞中观察到VDAC与GRP78以及VDAC与核糖体L28蛋白之间有显著水平的共定位。这是关于VDAC在JEV感染后重新定位的首次报道,并表明这可能是JEV在昆虫细胞中的复制策略的一部分。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1c91/4121193/64b539a2cdbe/TSWJ2014-976015.001.jpg

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