Abstract
BackgroundJapanese encephalitis virus (JEV) is a member of the mosquito-borne Flaviviridae family of viruses that causes human encephalitis. Upon infection of a new host, replication of viral RNA involves not only the viral RNA-dependent RNA polymerase (RdRp), but also host proteins. Host factors involved in JEV replication are not well characterized.ResultsWe identified Hdj2, a heat-shock protein 40 (Hsp40)/DnaJ homolog, from a mouse brain cDNA library interacting with JEV nonstructural protein 5 (NS5) encoding viral RdRp using yeast two-hybrid system. Specific interaction of Hdj2 with NS5 was confirmed by coimmunoprecipitation and colocalization in JEV-infected cells. Overexpression of Hdj2 in JEV-infected cells led to an increase of RNA synthesis, and the virus titer was elevated approximately 4.5- to 10-fold. Knocking down of Hdj2 by siRNA reduced the virus production significantly.ConclusionsWe conclude that Hdj2 directly associates with JEV NS5 and facilitates viral replication. This study is the first to demonstrate Hdj2 involved in JEV replication, providing insight into a potential therapeutic target and cell-based vaccine development of JEV infection.
Highlights
Japanese encephalitis virus (JEV) is a member of the mosquito-borne Flaviviridae family of viruses that causes human encephalitis
JEV nonstructural protein 5 (NS5) protein interacts with Hdj2 in vivo Hdj2 is among the candidates that interact with NS5 by yeast two-hybrid assay; specificity of the interaction requires verification to avoid a false positive result
These observations indicate that Hdj2 interacts with NS5 in JEV-infected cells
Summary
Japanese encephalitis virus (JEV) is a member of the mosquito-borne Flaviviridae family of viruses that causes human encephalitis. Host factors involved in JEV replication are not well characterized. Japanese encephalitis virus (JEV), a mosquito-borne flavivirus, is a leading cause of encephalitis in Eastern and Southern Asia. There are three very effective JEV human vaccines that have been used and have reduced the impact of JEV infection in Asian countries. These are the Chinese-developed SA14-14-2 live-attenuated vaccine, the conventional suckling mouse-brain derived, formalin-inactivated vaccine, and the Vero cell derived, formalin-inactivated vaccine [1,2]. The JEV genome is a single-strand positive sense RNA, 10,976 nucleotides (nts) in length.
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