Abstract
Replication of the Japanese encephalitis virus (JEV) genome depends on host factors for successfully completing their life cycles; to do this, host factors have been recruited and/or relocated to the site of viral replication. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), a cellular metabolic protein, was found to colocalize with viral RNA-dependent RNA polymerase (NS5) in JEV-infected cells. Subcellular fractionation further indicated that GAPDH remained relatively constant in the cytosol, while increasing at 12 to 24 hours postinfection (hpi) and decreasing at 36 hpi in the nuclear fraction of infected cells. In contrast, the redistribution patterns of GAPDH were not observed in the uninfected cells. Co-immunoprecipitation of GAPDH and JEV NS5 protein revealed no direct protein-protein interaction; instead, GAPDH binds to the 3' termini of plus- and minus-strand RNAs of JEV by electrophoretic mobility shift assays. Accordingly, GAPDH binds to the minus strand more efficiently than to the plus strand of JEV RNAs. This study highlights the findings that infection of JEV changes subcellular localization of GAPDH suggesting that this metabolic enzyme may play a role in JEV replication.
Highlights
Japanese encephalitis virus (JEV) is a mosquito-borne flavivirus that causes acute encephalitis in humans, with a high fatality rate of 20 to 50% [1]
We demonstrate that the colocalization of Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) with NS5 in JEV-infected cells was via binding to viral RNAs rather than binding to the NS5 protein directly
We have shown that GAPDH colocalized with JEV NS5 in the infected cells (Fig. 1), adding that this colocalization may be due to its ubiquitous nature and relatively high abundance
Summary
Japanese encephalitis virus (JEV) is a mosquito-borne flavivirus that causes acute encephalitis in humans, with a high fatality rate of 20 to 50% [1]. It contains a singlestranded positive RNA genome of 10,976 nucleotides (nts) in length that encodes a single large open reading frame (ORF) flanked by a 5'-untranslated region (5'-UTR, 95 nts) and a 3'-untranslated region (3'-UTR, 585 nts). It has been reported that the NS5 protein binds to the 3' end stemloop (SL) RNA as well as associates with NS3 in the replication complex (RC) [4]. The major replicase proteins NS3 and NS5 were found to localize within the nucleus [6,7]. The reason for nuclear localization of these viral proteins remains unknown; The cost of publication in Journal of Biomedical Science is bourne by the National Science Council,Taiwan
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