A conformation-based intra-molecular initiation factor identified in the flavivirus RNA-dependent RNA polymerase.
The flaviviruses pose serious threats to human health. Being a natural fusion of a methyltransferase (MTase) and an RNA-dependent RNA polymerase (RdRP), NS5 is the most conserved flavivirus protein and an important antiviral target. Previously reported NS5 structures represented by those from the Ja...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2020-05-01
|
| Series: | PLoS Pathogens |
| Online Access: | https://doi.org/10.1371/journal.ppat.1008484 |
| _version_ | 1818348755816022016 |
|---|---|
| author | Jiqin Wu Han-Qing Ye Qiu-Yan Zhang Guoliang Lu Bo Zhang Peng Gong |
| author_facet | Jiqin Wu Han-Qing Ye Qiu-Yan Zhang Guoliang Lu Bo Zhang Peng Gong |
| author_sort | Jiqin Wu |
| collection | DOAJ |
| description | The flaviviruses pose serious threats to human health. Being a natural fusion of a methyltransferase (MTase) and an RNA-dependent RNA polymerase (RdRP), NS5 is the most conserved flavivirus protein and an important antiviral target. Previously reported NS5 structures represented by those from the Japanese encephalitis virus (JEV) and Dengue virus serotype 3 (DENV3) exhibit two apparently different global conformations, defining two sets of intra-molecular MTase-RdRP interactions. However, whether these NS5 conformations are conserved in flaviviruses and their specific functions remain elusive. Here we report two forms of DENV serotype 2 (DENV2) NS5 crystal structures representing two conformational states with defined analogies to the JEV-mode and DENV3-mode conformations, respectively, demonstrating the conservation of both conformation modes and providing clues for how different conformational states may be interconnected. Data from in vitro polymerase assays further demonstrate that perturbing the JEV-mode but not the DENV3-mode intra-molecular interactions inhibits catalysis only at initiation, while the cell-based virological analysis suggests that both modes of interactions are important for virus proliferation. Our work highlights the role of MTase as a unique intra-molecular initiation factor specifically only through the JEV-mode conformation, providing an example of conformation-based crosstalk between naturally fused protein functional modules. |
| first_indexed | 2024-12-13T17:55:06Z |
| format | Article |
| id | doaj.art-0d0d0a1c1e33491aab7fb26012dcc83e |
| institution | Directory Open Access Journal |
| issn | 1553-7366 1553-7374 |
| language | English |
| last_indexed | 2024-12-13T17:55:06Z |
| publishDate | 2020-05-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Pathogens |
| spelling | doaj.art-0d0d0a1c1e33491aab7fb26012dcc83e2022-12-21T23:36:23ZengPublic Library of Science (PLoS)PLoS Pathogens1553-73661553-73742020-05-01165e100848410.1371/journal.ppat.1008484A conformation-based intra-molecular initiation factor identified in the flavivirus RNA-dependent RNA polymerase.Jiqin WuHan-Qing YeQiu-Yan ZhangGuoliang LuBo ZhangPeng GongThe flaviviruses pose serious threats to human health. Being a natural fusion of a methyltransferase (MTase) and an RNA-dependent RNA polymerase (RdRP), NS5 is the most conserved flavivirus protein and an important antiviral target. Previously reported NS5 structures represented by those from the Japanese encephalitis virus (JEV) and Dengue virus serotype 3 (DENV3) exhibit two apparently different global conformations, defining two sets of intra-molecular MTase-RdRP interactions. However, whether these NS5 conformations are conserved in flaviviruses and their specific functions remain elusive. Here we report two forms of DENV serotype 2 (DENV2) NS5 crystal structures representing two conformational states with defined analogies to the JEV-mode and DENV3-mode conformations, respectively, demonstrating the conservation of both conformation modes and providing clues for how different conformational states may be interconnected. Data from in vitro polymerase assays further demonstrate that perturbing the JEV-mode but not the DENV3-mode intra-molecular interactions inhibits catalysis only at initiation, while the cell-based virological analysis suggests that both modes of interactions are important for virus proliferation. Our work highlights the role of MTase as a unique intra-molecular initiation factor specifically only through the JEV-mode conformation, providing an example of conformation-based crosstalk between naturally fused protein functional modules.https://doi.org/10.1371/journal.ppat.1008484 |
| spellingShingle | Jiqin Wu Han-Qing Ye Qiu-Yan Zhang Guoliang Lu Bo Zhang Peng Gong A conformation-based intra-molecular initiation factor identified in the flavivirus RNA-dependent RNA polymerase. PLoS Pathogens |
| title | A conformation-based intra-molecular initiation factor identified in the flavivirus RNA-dependent RNA polymerase. |
| title_full | A conformation-based intra-molecular initiation factor identified in the flavivirus RNA-dependent RNA polymerase. |
| title_fullStr | A conformation-based intra-molecular initiation factor identified in the flavivirus RNA-dependent RNA polymerase. |
| title_full_unstemmed | A conformation-based intra-molecular initiation factor identified in the flavivirus RNA-dependent RNA polymerase. |
| title_short | A conformation-based intra-molecular initiation factor identified in the flavivirus RNA-dependent RNA polymerase. |
| title_sort | conformation based intra molecular initiation factor identified in the flavivirus rna dependent rna polymerase |
| url | https://doi.org/10.1371/journal.ppat.1008484 |
| work_keys_str_mv | AT jiqinwu aconformationbasedintramolecularinitiationfactoridentifiedintheflavivirusrnadependentrnapolymerase AT hanqingye aconformationbasedintramolecularinitiationfactoridentifiedintheflavivirusrnadependentrnapolymerase AT qiuyanzhang aconformationbasedintramolecularinitiationfactoridentifiedintheflavivirusrnadependentrnapolymerase AT guolianglu aconformationbasedintramolecularinitiationfactoridentifiedintheflavivirusrnadependentrnapolymerase AT bozhang aconformationbasedintramolecularinitiationfactoridentifiedintheflavivirusrnadependentrnapolymerase AT penggong aconformationbasedintramolecularinitiationfactoridentifiedintheflavivirusrnadependentrnapolymerase AT jiqinwu conformationbasedintramolecularinitiationfactoridentifiedintheflavivirusrnadependentrnapolymerase AT hanqingye conformationbasedintramolecularinitiationfactoridentifiedintheflavivirusrnadependentrnapolymerase AT qiuyanzhang conformationbasedintramolecularinitiationfactoridentifiedintheflavivirusrnadependentrnapolymerase AT guolianglu conformationbasedintramolecularinitiationfactoridentifiedintheflavivirusrnadependentrnapolymerase AT bozhang conformationbasedintramolecularinitiationfactoridentifiedintheflavivirusrnadependentrnapolymerase AT penggong conformationbasedintramolecularinitiationfactoridentifiedintheflavivirusrnadependentrnapolymerase |