Japanese encephalitis virus co-opts the ER-stress response protein GRP78 for viral infectivity.
Japanese encephalitis virus co-opts the ER-stress response protein GRP78 for viral infectivity.
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DOI：
10.1186/1743-422x-8-128
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发表时间：
2011-03-20
期刊：
影响因子：
4.8
通讯作者：
Wang RY
中科院分区：
文献类型：
--
作者：
Wu YP;Chang CM;Hung CY;Tsai MC;Schuyler SC;Wang RY
通过液相色谱串联质谱法（LC-MS）分析来自日本脑炎病毒（JEV）感染的婴儿肾脏-21（BHK-21）细胞培养物的无血清培养基，以鉴定病毒感染后分泌的宿主蛋白。鉴定了五种蛋白质，包括分子伴侣Hsp 90、GRP 78和Hsp 70。然后研究GRP 78在JEV生命周期中的功能作用。在蔗糖密度梯度中观察到GRP 78与JEV颗粒的共迁移，并通过免疫荧光分析检测到病毒E蛋白与GRP 78的共定位。通过siRNA敲低GRP 78表达不影响病毒RNA复制，但确实损害成熟病毒的产生。不与GPR 78共裂解的成熟病毒显示病毒感染性显著降低。我们的研究结果支持的假设，即JEV coo-opts宿主细胞GPR 78用于病毒成熟和随后的细胞感染。
The serum-free medium from Japanese encephalitis virus (JEV) infected Baby Hamster Kidney-21 (BHK-21) cell cultures was analyzed by liquid chromatography tandem mass spectrometry (LC-MS) to identify host proteins that were secreted upon viral infection. Five proteins were identified, including the molecular chaperones Hsp90, GRP78, and Hsp70. The functional role of GRP78 in the JEV life cycle was then investigated. Co-migration of GRP78 with JEV particles in sucrose density gradients was observed and co-localization of viral E protein with GRP78 was detected by immunofluorescence analysis in vivo. Knockdown of GRP78 expression by siRNA did not effect viral RNA replication, but did impair mature viral production. Mature viruses that do not co-fractionate with GPR78 displayed a significant decrease in viral infectivity. Our results support the hypothesis that JEV co-opts host cell GPR78 for use in viral maturation and in subsequent cellular infections.