Structural insights into RNA recognition by the Chikungunya virus nsP2 helicase

doi:10.1073/pnas.1900656116

	Structural insights into RNA recognition by the Chikungunya virus nsP2 helicase
	Law, Yee-Song 4,5; Utt, Age 6; Tan, Yaw Bia 4,5; Zheng, Jie 1,2; Wang, Sainan 6; Chen, Ming Wei 3,5; Griffin, Patrick R.1; Merits, Andres 6; Luo, Dahai 4,5
刊名	PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
	2019-05-07
卷号	116 期号:19 页码:9558-9567
关键词	Chikungunya virus nonstructural protein 2 superfamily 1 helicase ATP hydrolysis Alphavirus
ISSN号	0027-8424
DOI	10.1073/pnas.1900656116
通讯作者	Merits, Andres(andres.merits@ut.ee) ; Luo, Dahai(luodahai@ntu.edu.sg)
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 transitionstate 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.
资助项目	Ministry of Education of Singapore[MOE2016T22097] ; Ministry of Health of Singapore[NMRC OFIRG17nov084] ; Estonian Research Council[IUT 20-27]
WOS关键词	FUNCTIONAL INSIGHTS ; CRYSTAL-STRUCTURE ; SINDBIS VIRUS ; OLD-WORLD ; PROTEINS ; ALPHAVIRUSES ; REPLICATION ; COMPLEXES ; MECHANISM ; EXCHANGE
WOS研究方向	Science & Technology - Other Topics
语种	英语
出版者	NATL ACAD SCIENCES
WOS记录号	WOS:000467226400058
内容类型	期刊论文
源URL	[http://119.78.100.183/handle/2S10ELR8/289876]
专题	中国科学院上海药物研究所
通讯作者	Merits, Andres; Luo, Dahai
作者单位	1.Scripps Res Inst, Dept Mol Med, Jupiter, FL 33458 USA 2.Chinese Acad Sci, Shanghai Inst Mat Med, Shanghai 201203, Peoples R China 3.Nanyang Technol Univ, Sch Biol Sci, Singapore 637551, Singapore 4.Nanyang Technol Univ, Lee Kong Chian Sch Med, Singapore 636921, Singapore 5.Nanyang Technol Univ, NTU Inst Struct Biol, Singapore 636921, Singapore 6.Univ Tartu, Inst Technol, EE-50411 Tartu, Estonia
推荐引用方式 GB/T 7714	Law, Yee-Song,Utt, Age,Tan, Yaw Bia,et al. Structural insights into RNA recognition by the Chikungunya virus nsP2 helicase[J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA,2019,116(19):9558-9567.
APA	Law, Yee-Song.,Utt, Age.,Tan, Yaw Bia.,Zheng, Jie.,Wang, Sainan.,...&Luo, Dahai.(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),9558-9567.
MLA	Law, Yee-Song,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):9558-9567.