Molecular Function Of Hippo Signaling Pathway In Litopenaeus Vannamei During White Spot Syndrome Virus Infection

Litopenaeus vannamei is the most widely cultured and highest productive shrimp species in the world.However,the shrimp aquaculture is seriously threatened by many pathogens,especially white spot syndrome virus(WSSV),which is high infective and lethal to penaeus.Thus,improving knowledges of the interaction between WSSV and shrimp is necessary for promoting disease control as well.To date,many key molecules and cellular signaling pathways have been reported to be involved in WSSV-host interaction.Unfortunately,the studies in this area are scattered.In the present study,we used Cherax quadricarinatus Hpt cell to screen the compounds involved in early stage of WSSV-host interaction in large scale.Combining analysis of viral infection and cell toxicity test,we got 30 compounds whose targets were related to WSSV early infection.The targets of the compounds were involved in NF-κB signaling pathway,ion regulation,Hedgehog signaling pathway,retinoic acid signaling pathway and so on.According to the analysis of the targets of the compounds,it is suggestes that NFκB signaling pathway,Wnt signaling pathway and PI3K-Akt signaling pathway were all have close relationship with Hippo signaling pathway.Base on the previous work in the lab,Hippo signaling pathway was selected for further investigated.We cloned and identified the members of Hippo kinase cascade in L.vannamei,and named them lvhpo,lvsav,lvwts and lvmats,respectively.Sequence and phylogenetic analysis indicated Hippo kinase cascade is conserved in L.vannamei.Post WSSV infection,it was found that the transcriptional profiles of the members of Hippo kinase cascade were upregulated in hepatopancreas,while down-regulated in gills.This result indicated the tightly relatationship between Hippo signaling pathway with WSSV infection,and showed the different regulation in different tissues.Furthermore,we chose the key molecules LvHpo for investigation.The results showed that protein level of LvHpo were also decreased in shrimp gill during WSSV infection.Combining of knock-down of Ivhpo in vivo and overexpression of LvHpo in vitro indicated that LvHpo can regulate cell activity and apoptosis functioned on WSSV infection.Moreover,a comprehensive analysis of comparative transcriptome was performed to reveal the molecular mechanism and biological function of LvHpo.The result indicated LvHpo might be involved in the interaction between WSSV and host by regulation of AMPs,apoptosis and proliferation.In conclusion,this study screened the molecules inloved in WSSV and shrimp interaction in large-scale.Furthermore,the function of Hippo signaling pathway during WSSV infection was investigated.The study expanded the understanding of the interaction between WSSV and host,and provided beneficial enlightenment for promoting disease control.