Abstract
BackgroundThe JEV genome is a positive-sense RNA with a highly structured capped 5′UTR, 3′UTR and a large open reading frame. 3′UTR is the untranslated region of flavivirus and has various important functions during viral replication, such as translation, replication and encapsidation. During viral replication, the 3′UTR interacts with viral proteins and host proteins and is required for viral RNA replication and translocation.MethodsThe expression level of FUBP3 was knocked down by siRNA and Flag-tagged FUBP3 overexpression plasmid was constructed for overexpression. BHK-21 cells were cultured and infected with JEV to investigate the functional role of FUBP3 in the viral infection cycle. Subcellular localization of FUBP3 and viral replication complexes was observed by dual immunofluorescence staining.ResultsFour host proteins were specifically associated with the 3′UTR of JEV, and FUBP3 was selected to further investigate its potential functional role in the JEV infection cycle. Knockdown of FUBP3 protein resulted in a significant decrease in JEV viral titer, whereas ectopic overexpression of FUBP3 resulted in increased JE viral infectivity. In cells stably knocked down for FUBP3 and then infected with JEV, we found almost no detectable viral NS5 protein. In contrast, when cells stably knocking-down of FUBP3 overexpressed FUBP3, we found a significant increase in viral RNA production over time compared to controls. We also demonstrated that FUBP3 re-localized in the cytoplasm after infection with JEV and co-localized with viral proteins. Exogenous overexpression of FUBP3 was also shown to be located in the JE replication complex and to assist viral replication after JEV infection.ConclusionsThe overall results suggest that FUBP3 regulates RNA replication of JEV and promotes subsequent viral translation and viral particle production.
Highlights
Japanese encephalitis (JE) is an infectious disease caused by a Japanese encephalitis virus (JEV) that infects the central nervous system of humans
We identified four host proteins specificities associated with the JEV 3′Untranslated region (UTR), and FUBP3 and FUBP1 were among them
The FUBP3 protein interacted with the JEV 3′UTRTo identify host proteins bound to the JEV 3′ untranslated region (3′UTR), the RNA fragment was labeled with biotin and incubated with BHK-21 cell lysates
Summary
Japanese encephalitis (JE) is an infectious disease caused by a Japanese encephalitis virus (JEV) that infects the central nervous system of humans. The 3′UTR is the untranslated region of flaviviruses and has various important functions during viral replication, such as translation, replication and encapsidation [7, 8]. This 3′UTR contains several functional motifs, including conserved sequences (CS motifs), cyclization motifs, pseudoknot structure and 3′-stemloop (3′-SL) motif. In addition to RNA–RNA interactions within the viral genome, the 3′UTR of JEV interacts with viral proteins and host proteins that are required for viral RNA replication and translocation [14]. 3′UTR is the untranslated region of flavivirus and has various important functions during viral replication, such as translation, replication and encapsidation. The 3′UTR interacts with viral proteins and host proteins and is required for viral RNA replication and translocation
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
