Structural and Functional Studies of Chikungunya Virus nsP2
Chikungunya virus (CHIKV) is transmitted to humans through mosquitoes and causes Chikungunya fever. Nonstructural protein 2 (nsP2) contains an N-terminal RNA helicase with both nucleotide triphosphatase and RNA triphosphatase activities, and a C-terminal cysteine protease that is responsible for pol...
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MDPI AG
2020-07-01
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| Online Access: | https://www.mdpi.com/2504-3900/50/1/113 |
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| author | Yee Song Law Yaw Bia Tan Orion Shih Age Utt Jie Zheng Sainan Wang Ming Wei Chen Patrick R. Griffin U-Ser Jeng Andres Merits Dahai Luo |
| author_facet | Yee Song Law Yaw Bia Tan Orion Shih Age Utt Jie Zheng Sainan Wang Ming Wei Chen Patrick R. Griffin U-Ser Jeng Andres Merits Dahai Luo |
| author_sort | Yee Song Law |
| collection | DOAJ |
| description | Chikungunya virus (CHIKV) is transmitted to humans through mosquitoes and causes Chikungunya fever. Nonstructural protein 2 (nsP2) contains an N-terminal RNA helicase with both nucleotide triphosphatase and RNA triphosphatase activities, and a C-terminal cysteine protease that is responsible for polyprotein processing. Both N-terminal RNA helicase and C-terminal cysteine protease are connected through a flexible linker. Although the structure of the C-terminal cysteine protease has been solved, the structure and the conformational arrangement of full-length nsP2 remains elusive. Here, we determined the crystal structure of the helicase part of the CHIKV nsP2 (nsP2h) bound to the conserved 3′-end of the genomic RNA and the nucleotide analogue ADP-AlF<sub>4</sub>. The structure of this ternary complex revealed the molecular basis for viral RNA recognition and ATP hydrolysis by the nsP2h. Unique hydrophobic protein–RNA interactions play essential roles in viral RNA replication. We also determined the solution structure of full-length nsP2 using small-angle X-ray scattering (SAXS). The solution architecture of the nsP2 was modeled using the available high-resolution structures and program CORAL (complexes with random loops). The CORAL model revealed that nsP2 is partially unfolded and the N-terminal protease domain is arranged near the N-terminal domain of the helicase domain. These findings expand our knowledge of CHIKV and related alphaviruses and might also have broad implications for antiviral and vaccine developments against pathogenic alphaviruses. |
| first_indexed | 2024-03-10T18:45:55Z |
| format | Article |
| id | doaj.art-dd0e57e9f5c049399e471f1fc3668f60 |
| institution | Directory Open Access Journal |
| issn | 2504-3900 |
| language | English |
| last_indexed | 2025-03-20T05:50:32Z |
| publishDate | 2020-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Proceedings |
| spelling | doaj.art-dd0e57e9f5c049399e471f1fc3668f602024-10-03T02:28:51ZengMDPI AGProceedings2504-39002020-07-0150111310.3390/proceedings2020050113Structural and Functional Studies of Chikungunya Virus nsP2Yee Song Law0Yaw Bia Tan1Orion Shih2Age Utt3Jie Zheng4Sainan Wang5Ming Wei Chen6Patrick R. Griffin7U-Ser Jeng8Andres Merits9Dahai Luo10Lee Kong Chian School of Medicine, Nanyang Technological University, EMB 03-07, 59 Nanyang Drive, Singapore 636921, SingaporeLee Kong Chian School of Medicine, Nanyang Technological University, EMB 03-07, 59 Nanyang Drive, Singapore 636921, SingaporeNational Synchrotron Radiation Research Center, Hsinchu 30076, TaiwanInstitute of Technology, University of Tartu, Tartu 50411, EstoniaDepartment of Molecular Medicine, The Scripps Research Institute, Jupiter, FL 33458, USAInstitute of Technology, University of Tartu, Tartu 50411, EstoniaNTU Institute of Structural Biology, Nanyang Technological University, EMB 06-01, 59 Nanyang Drive, Singapore 636921, SingaporeDepartment of Molecular Medicine, The Scripps Research Institute, Jupiter, FL 33458, USANational Synchrotron Radiation Research Center, Hsinchu 30076, TaiwanInstitute of Technology, University of Tartu, Tartu 50411, EstoniaLee Kong Chian School of Medicine, Nanyang Technological University, EMB 03-07, 59 Nanyang Drive, Singapore 636921, SingaporeChikungunya virus (CHIKV) is transmitted to humans through mosquitoes and causes Chikungunya fever. Nonstructural protein 2 (nsP2) contains an N-terminal RNA helicase with both nucleotide triphosphatase and RNA triphosphatase activities, and a C-terminal cysteine protease that is responsible for polyprotein processing. Both N-terminal RNA helicase and C-terminal cysteine protease are connected through a flexible linker. Although the structure of the C-terminal cysteine protease has been solved, the structure and the conformational arrangement of full-length nsP2 remains elusive. Here, we determined the crystal structure of the helicase part of the CHIKV nsP2 (nsP2h) bound to the conserved 3′-end of the genomic RNA and the nucleotide analogue ADP-AlF<sub>4</sub>. The structure of this ternary complex revealed the molecular basis for viral RNA recognition and ATP hydrolysis by the nsP2h. Unique hydrophobic protein–RNA interactions play essential roles in viral RNA replication. We also determined the solution structure of full-length nsP2 using small-angle X-ray scattering (SAXS). The solution architecture of the nsP2 was modeled using the available high-resolution structures and program CORAL (complexes with random loops). The CORAL model revealed that nsP2 is partially unfolded and the N-terminal protease domain is arranged near the N-terminal domain of the helicase domain. These findings expand our knowledge of CHIKV and related alphaviruses and might also have broad implications for antiviral and vaccine developments against pathogenic alphaviruses.https://www.mdpi.com/2504-3900/50/1/113Chikungunya virushelicaseprotease |
| spellingShingle | Yee Song Law Yaw Bia Tan Orion Shih Age Utt Jie Zheng Sainan Wang Ming Wei Chen Patrick R. Griffin U-Ser Jeng Andres Merits Dahai Luo Structural and Functional Studies of Chikungunya Virus nsP2 Proceedings Chikungunya virus helicase protease |
| title | Structural and Functional Studies of Chikungunya Virus nsP2 |
| title_full | Structural and Functional Studies of Chikungunya Virus nsP2 |
| title_fullStr | Structural and Functional Studies of Chikungunya Virus nsP2 |
| title_full_unstemmed | Structural and Functional Studies of Chikungunya Virus nsP2 |
| title_short | Structural and Functional Studies of Chikungunya Virus nsP2 |
| title_sort | structural and functional studies of chikungunya virus nsp2 |
| topic | Chikungunya virus helicase protease |
| url | https://www.mdpi.com/2504-3900/50/1/113 |
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