Transcriptomic response of mosquitoes to Japanese encephalitis virus and identification of its potential entry factors.

Japanese encephalitis virus (JEV), an Orthoflavivirus transmitted by mosquitoes, can cause severe disease in humans, yet its interactions with mosquito vectors remain poorly understood. This study presents the first de novo assembled transcriptome of Culex annulirostris, a major Australian JEV vector, and examines its transcriptional response to infection with JEV. A total of 350, 322, and 451 genes were differentially expressed at 2, 7, and 12 days post-infection, respectively, involving functions related to membrane-bounded organelles, protein metabolic processes, and catalytic activity. To identify potential viral entry factors, JEV Envelope protein was incubated with lysates from Culex quinquefasciatus Hsu cells. Co-immunoprecipitation and mass spectrometry revealed prohibitin, annexin, and 14-3-3ε as interacting proteins. Silencing these genes reduced JEV infection, and anti-prohibitin antibodies further suppressed viral replication. These findings highlight mosquito transcriptomic responses to JEV and identify annexin, 14-3-3ε, and prohibitin-especially prohibitin-as pro-viral factors and potential targets for blocking JEV entry.