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PMID:24086464
| Citation |
Ye, J, Chen, Z, Zhang, B, Miao, H, Zohaib, A, Xu, Q, Chen, H and Cao, S (2013) Heat Shock Protein 70 Is Associated with Replicase Complex of Japanese Encephalitis Virus and Positively Regulates Viral Genome Replication. PLoS ONE 8:e75188 |
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| Abstract |
Japanese encephalitis virus (JEV) is a mosquito-borne flavivirus that causes the most prevalent viral encephalitis in Asia. The NS5 protein of JEV is a key component of the viral replicase complex, which plays a crucial role in viral pathogenesis. In this study, tandem affinity purification (TAP) followed by mass spectrometry analysis was performed to identify novel host proteins that interact with NS5. Heat shock protein 70 (Hsp70), eukaryotic elongation factor 1-alpha (eEF-1α) and ras-related nuclear protein (Ran) were demonstrated to interact with NS5. In addition to NS5, Hsp70 was also found to interact with NS3 which is another important member of the replicase complex. It was observed that the cytoplasmic Hsp70 partially colocalizes with the components of viral replicase complex including NS3, NS5 and viral dsRNA during JEV infection. Knockdown of Hsp70 resulted in a significantly reduced JEV genome replication. Further analysis reveals that Hsp70 enhances the stability of viral proteins in JEV replicase complex. These results suggest an important role for Hsp70 in regulating JEV replication, which provides a potential target for the development of anti-JEV therapies. |
| Links |
PubMed PMC3781048 Online version:10.1371/journal.pone.0075188 |
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Significance
Annotations
| Gene product | Qualifier | GO Term | Evidence Code | with/from | Aspect | Extension | Notes | Status |
|---|---|---|---|---|---|---|---|---|
| GO:0016563: transcription activator activity |
ECO:0000314: |
F |
Fig.7 In vitro transcription analyses. |
complete | ||||
| GO:0008166: viral replication |
ECO:0000315: |
P |
Figure 3C.The purified proteins of GST-NS5 (406-905) and His-Hsp70 were analyzed by SDS-PAGE. |
complete | ||||
| GO:0008166: |
ECO:0000314: |
Figure 4A. Interaction of Hsp70 with components of the JEV replicase complex. Interaction of Hsp70 with JEV NS3. 293T cells were co-transfected with Myc-tagged Hsp70 plasmid and Flag-tagged NS3 plasmid or vector. Cell lysates were harvested 36 h after transfection for co-IP experiments with anti-Flag antibody. The precipitates were then analyzed by Western blotting with anti-Flag or anti-Myc antibody. |
complete | |||||
| GO:0008166: viral replication |
ECO:0000314: |
P |
Figure 4 (B) Association of Hsp70 with NS3 and NS5 proteins and viral RNA in JEV-infected cells. 293T cells were transfected with Hsp70-Myc expressing plasmid or vector before being mock-infected or infected with JEV at an MOI of 1.0. Cell lysates were harvested at 36 h p.i. and subjected to immunoprecipitation with antibody against Myc, followed by Western blotting for the detection of NS3 and NS5. The RNA were extracted from the precipitates and then subjected to RT-PCR for detecting viral RNA. RNA extracted from cells infected and uninfected with JEV was used as positive control (PC) and negative control (NC) respectively. |
complete | ||||
See also
References
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