Hsp40 Protein DNAJB6 Interacts with Viral NS3 and Inhibits the Replication of the Japanese Encephalitis Virus
Hsp40 Protein DNAJB6 Interacts with Viral NS3 and Inhibits the Replication of the Japanese Encephalitis Virus
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DOI：
10.3390/ijms20225719
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发表时间：
2019-11-01
影响因子：
5.6
通讯作者：
Cao, San-Jie
中科院分区：
文献类型：
--
作者：
Cao, Yu-Qin;Yuan, Lei;Cao, San-Jie
日本脑炎病毒（JEV）是一种蚊媒黄病毒，流行于东亚和东南亚、西太平洋和澳大利亚北方。由于病毒是专性细胞内病原体，病毒进入，复制和组装的动态过程依赖于许多宿主-病原体相互作用。目前正在努力确定与JEV相互作用的宿主因素，因为它们的鉴定和表征仍然不完整。黄病毒非结构蛋白3（NS 3）的三种酶活性，包括丝氨酸蛋白酶、RNA解旋酶和三磷酸酶，在黄病毒的生命周期中起主要作用。为了鉴定与NS 3相互作用的细胞因子，我们使用酵母双杂交测定筛选人脑cDNA文库，并鉴定出8种与NS 3相互作用的蛋白质：COPS 5、FBLN 5、PPP 2CB、CRBN、DNAJB 6、UBE 2N、ZNF 350和GPR 137 B。我们证明了DnaJ热休克蛋白家族（Hsp 40）成员B6（DNAJB 6）与NS 3共定位并相互作用，并在JEV复制中具有负调控功能。我们还表明，DNAJB 6功能的丧失导致病毒复制显著增加，但不影响病毒结合或内化。此外，JEV感染期间DNAJB 6破坏的时间过程以病毒载量依赖性方式变化，这表明JEV靶向这种宿主伴侣蛋白以获得病毒益处。破译NS 3相互作用宿主蛋白在病毒体产生中的功能模式将揭示JEV致病机制，也可能揭示抗病毒治疗的新途径。
The Japanese encephalitis virus (JEV) is a mosquito-borne flavivirus prevalent in east and southeast Asia, the Western Pacific, and northern Australia. Since viruses are obligatory intracellular pathogens, the dynamic processes of viral entry, replication, and assembly are dependent on numerous host-pathogen interactions. Efforts to identify JEV-interacting host factors are ongoing because their identification and characterization remain incomplete. Three enzymatic activities of flavivirus non-structural protein 3 (NS3), including serine protease, RNA helicase, and triphosphatase, play major roles in the flaviviruses lifecycle. To identify cellular factors that interact with NS3, we screened a human brain cDNA library using a yeast two-hybrid assay, and identified eight proteins that putatively interact with NS3: COPS5, FBLN5, PPP2CB, CRBN, DNAJB6, UBE2N, ZNF350, and GPR137B. We demonstrated that the DnaJ heat shock protein family (Hsp40) member B6 (DNAJB6) colocalizes and interacts with NS3, and has a negative regulatory function in JEV replication. We also show that loss of DNAJB6 function results in significantly increased viral replication, but does not affect viral binding or internalization. Moreover, the time-course of DNAJB6 disruption during JEV infection varies in a viral load-dependent manner, suggesting that JEV targets this host chaperone protein for viral benefit. Deciphering the modes of NS3-interacting host proteins functions in virion production will shed light on JEV pathogenic mechanisms and may also reveal new avenues for antiviral therapeutics.