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DOI	10.1073/pnas.1900656116
	Structural insights into RNA recognition by the Chikungunya virus nsP2 helicase
	Law Y.-S.; Utt A.; Tan Y.B.; Zheng J.; Wang S.; Chen M.W.; Griffin P.R.; Merits A.; Luo D.
发表日期	2019
ISSN	0027-8424
起始页码	9558
结束页码	9567
卷号	116 期号:19
英文摘要	Chikungunya virus (CHIKV) is transmitted to humans through mosquitoes and causes Chikungunya fever. Nonstructural protein 2 (nsP2) exhibits the protease and RNA helicase activities that are required for viral RNA replication and transcription. Unlike for the C-terminal protease, the structure of the N-terminal RNA helicase (nsP2h) has not been determined. Here, we report the crystal structure of the nsP2h bound to the conserved 3′-end 14 nucleotides of the CHIKV genome and the nonhydrolyzable transition-state nucleotide analog ADP-AlF4. Overall, the structural analysis revealed that nsP2h adopts a uniquely folded N-terminal domain followed by a superfamily 1 RNA helicase fold. The conserved helicase motifs establish polar contacts with the RNA backbone. There are three hydrophobic residues (Y161, F164, and F287) which form stacking interactions with RNA bases and thereby bend the RNA backbone. An F287A substitution that disrupted these stacking interactions increased the basal ATPase activity but decreased the RNA binding affinity. Furthermore, the F287A substitution reduced viral infectivity by attenuating subgenomic RNA synthesis. Replication of the mutant virus was restored by pseudoreversion (A287V) or adaptive mutations in the RecA2 helicase domain (T358S or V410I). Y161A and/or F164A substitutions, which were designed to disrupt the interactions with the RNA molecule, did not affect the ATPase activity but completely abolished the replication and transcription of viral RNA and the infectivity of CHIKV. Our study sheds light on the roles of the RNA helicase region in viral replication and provides insights that might be applicable to alphaviruses and other RNA viruses in general. © 2019 National Academy of Sciences. All rights reserved.
英文关键词	Alphavirus; ATP hydrolysis; Chikungunya virus; Nonstructural protein 2; Superfamily 1 helicase
语种	英语
scopus关键词	adenosine diphosphate; adenosine triphosphatase; RNA helicase; adenosine diphosphate tetrafluoroaluminate; organometallic compound; RNA helicase; viral protein; virus RNA; Alphavirus; amino acid substitution; amino terminal sequence; Article; binding affinity; Chikungunya virus; controlled study; crystal structure; enzyme activity; enzyme structure; gene mutation; hydrophobicity; nonhuman; priority journal; protein binding; protein family; protein folding; protein interaction; protein motif; RNA binding; RNA recognition motif; RNA synthesis; RNA transcription; RNA virus; structure analysis; U2OS cell line; virus genome; virus replication; virus virulence; biosynthesis; chemistry; Chikungunya virus; metabolism; protein domain; Adenosine Diphosphate; Chikungunya virus; Organometallic Compounds; Protein Domains; RNA Helicases; RNA, Viral; Viral Proteins
来源期刊	Proceedings of the National Academy of Sciences of the United States of America
文献类型	期刊论文
条目标识符	http://gcip.llas.ac.cn/handle/2XKMVOVA/160368
作者单位	Law, Y.-S., Lee Kong Chian School of Medicine, Nanyang Technological University, Singapore, 636921, Singapore, NTU Institute of Structural Biology, Nanyang Technological University, Singapore, 636921, Singapore; Utt, A., Institute of Technology, University of Tartu, Tartu, 50411, Estonia; Tan, Y.B., Lee Kong Chian School of Medicine, Nanyang Technological University, Singapore, 636921, Singapore, NTU Institute of Structural Biology, Nanyang Technological University, Singapore, 636921, Singapore; Zheng, J., Department of Molecular Medicine, Scripps Research Institute, Jupiter, FL 33458, United States, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, 201203, China; Wang, S., Institute of Technology, University of Tartu, Tartu, 50411, Estonia; Chen, M.W., NTU Institute of Structural Biology, Nanyang Technological University, Singapore, 636921, Singapore, School of Biological Sciences, Nanyang Technological University, Singapore, 637551, Singapore; Griffin, P.R., Department of Mol...
推荐引用方式 GB/T 7714	Law Y.-S.,Utt A.,Tan Y.B.,et al. Structural insights into RNA recognition by the Chikungunya virus nsP2 helicase[J],2019,116(19).
APA	Law Y.-S..,Utt A..,Tan Y.B..,Zheng J..,Wang S..,...&Luo D..(2019).Structural insights into RNA recognition by the Chikungunya virus nsP2 helicase.Proceedings of the National Academy of Sciences of the United States of America,116(19).
MLA	Law Y.-S.,et al."Structural insights into RNA recognition by the Chikungunya virus nsP2 helicase".Proceedings of the National Academy of Sciences of the United States of America 116.19(2019).