Crystal structures of alphavirus nonstructural protein 4 (nsP4) reveal an intrinsically dynamic RNA-dependent RNA polymerase fold
Alphaviruses such as Ross River virus (RRV), chikungunya virus (CHIKV), Sindbis virus (SINV), and Venezuelan equine encephalitis virus (VEEV) are mosquito-borne pathogens that can cause arthritis or encephalitis diseases. Nonstructural protein 4 (nsP4) of alphaviruses possesses RNA-dependent RNA pol...
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| Format: | Journal Article |
| Language: | English |
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2022
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| Online Access: | https://hdl.handle.net/10356/161300 |
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| author | Tan, Yaw Bia Lello, Laura Sandra Liu, Xin Law, Yee-Song Kang, Congbao Lescar, Julien Zheng, Jie Merits, Andres Luo, Dahai |
| author2 | Lee Kong Chian School of Medicine (LKCMedicine) |
| author_facet | Lee Kong Chian School of Medicine (LKCMedicine) Tan, Yaw Bia Lello, Laura Sandra Liu, Xin Law, Yee-Song Kang, Congbao Lescar, Julien Zheng, Jie Merits, Andres Luo, Dahai |
| author_sort | Tan, Yaw Bia |
| collection | NTU |
| description | Alphaviruses such as Ross River virus (RRV), chikungunya virus (CHIKV), Sindbis virus (SINV), and Venezuelan equine encephalitis virus (VEEV) are mosquito-borne pathogens that can cause arthritis or encephalitis diseases. Nonstructural protein 4 (nsP4) of alphaviruses possesses RNA-dependent RNA polymerase (RdRp) activity essential for viral RNA replication. No 3D structure has been available for nsP4 of any alphaviruses despite its importance for understanding alphaviral RNA replication and for the design of antiviral drugs. Here, we report crystal structures of the RdRp domain of nsP4 from both RRV and SINV determined at resolutions of 2.6 Å and 1.9 Å. The structure of the alphavirus RdRp domain appears most closely related to RdRps from pestiviruses, noroviruses, and picornaviruses. Hydrogen-deuterium exchange mass spectrometry (HDX-MS) and nuclear magnetic resonance (NMR) methods showed that in solution, nsP4 is highly dynamic with an intrinsically disordered N-terminal domain. Both full-length nsP4 and the RdRp domain were capable to catalyze RNA polymerization. Structure-guided mutagenesis using a trans-replicase system identified nsP4 regions critical for viral RNA replication. |
| first_indexed | 2024-10-01T06:53:57Z |
| format | Journal Article |
| id | ntu-10356/161300 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T06:53:57Z |
| publishDate | 2022 |
| record_format | dspace |
| spelling | ntu-10356/1613002023-02-28T17:13:55Z Crystal structures of alphavirus nonstructural protein 4 (nsP4) reveal an intrinsically dynamic RNA-dependent RNA polymerase fold Tan, Yaw Bia Lello, Laura Sandra Liu, Xin Law, Yee-Song Kang, Congbao Lescar, Julien Zheng, Jie Merits, Andres Luo, Dahai Lee Kong Chian School of Medicine (LKCMedicine) School of Biological Sciences NTU Institute of Structural Biology Science::Medicine Science::Biological sciences Viral Protein Virus RNA Alphaviruses such as Ross River virus (RRV), chikungunya virus (CHIKV), Sindbis virus (SINV), and Venezuelan equine encephalitis virus (VEEV) are mosquito-borne pathogens that can cause arthritis or encephalitis diseases. Nonstructural protein 4 (nsP4) of alphaviruses possesses RNA-dependent RNA polymerase (RdRp) activity essential for viral RNA replication. No 3D structure has been available for nsP4 of any alphaviruses despite its importance for understanding alphaviral RNA replication and for the design of antiviral drugs. Here, we report crystal structures of the RdRp domain of nsP4 from both RRV and SINV determined at resolutions of 2.6 Å and 1.9 Å. The structure of the alphavirus RdRp domain appears most closely related to RdRps from pestiviruses, noroviruses, and picornaviruses. Hydrogen-deuterium exchange mass spectrometry (HDX-MS) and nuclear magnetic resonance (NMR) methods showed that in solution, nsP4 is highly dynamic with an intrinsically disordered N-terminal domain. Both full-length nsP4 and the RdRp domain were capable to catalyze RNA polymerization. Structure-guided mutagenesis using a trans-replicase system identified nsP4 regions critical for viral RNA replication. Ministry of Education (MOE) Published version Singapore Ministry of Education under its Singapore Ministry of Education Academic Research Fund Tier 2 [MOET2EP30220-0009, MOE2016T22097]; Estonian Research Council [PRG1154]. Funding for open access charge: Ministry of Education of Singapore [MOE2016T22097]. 2022-08-24T05:32:07Z 2022-08-24T05:32:07Z 2022 Journal Article Tan, Y. B., Lello, L. S., Liu, X., Law, Y., Kang, C., Lescar, J., Zheng, J., Merits, A. & Luo, D. (2022). Crystal structures of alphavirus nonstructural protein 4 (nsP4) reveal an intrinsically dynamic RNA-dependent RNA polymerase fold. Nucleic Acids Research, 50(2), 1000-1016. https://dx.doi.org/10.1093/nar/gkab1302 0305-1048 https://hdl.handle.net/10356/161300 10.1093/nar/gkab1302 35037043 2-s2.0-85125005502 2 50 1000 1016 en MOET2EP30220-0009 MOE2016T22097 Nucleic Acids Research © The Author(s) 2022. Published by Oxford University Press on behalf of Nucleic Acids Research. This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com application/pdf |
| spellingShingle | Science::Medicine Science::Biological sciences Viral Protein Virus RNA Tan, Yaw Bia Lello, Laura Sandra Liu, Xin Law, Yee-Song Kang, Congbao Lescar, Julien Zheng, Jie Merits, Andres Luo, Dahai Crystal structures of alphavirus nonstructural protein 4 (nsP4) reveal an intrinsically dynamic RNA-dependent RNA polymerase fold |
| title | Crystal structures of alphavirus nonstructural protein 4 (nsP4) reveal an intrinsically dynamic RNA-dependent RNA polymerase fold |
| title_full | Crystal structures of alphavirus nonstructural protein 4 (nsP4) reveal an intrinsically dynamic RNA-dependent RNA polymerase fold |
| title_fullStr | Crystal structures of alphavirus nonstructural protein 4 (nsP4) reveal an intrinsically dynamic RNA-dependent RNA polymerase fold |
| title_full_unstemmed | Crystal structures of alphavirus nonstructural protein 4 (nsP4) reveal an intrinsically dynamic RNA-dependent RNA polymerase fold |
| title_short | Crystal structures of alphavirus nonstructural protein 4 (nsP4) reveal an intrinsically dynamic RNA-dependent RNA polymerase fold |
| title_sort | crystal structures of alphavirus nonstructural protein 4 nsp4 reveal an intrinsically dynamic rna dependent rna polymerase fold |
| topic | Science::Medicine Science::Biological sciences Viral Protein Virus RNA |
| url | https://hdl.handle.net/10356/161300 |
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