Effect of significant variation in nonstructural protein 1 of 2019-nCoV on its binding ability to viral 5′UTR
10.3760/cma.j.cn112866-20230427-00044
- VernacularTitle:新型冠状病毒非结构蛋白1重要变异对其与病毒5’UTR结合能力的影响
- Author:
Qiao QIAO
1
;
Xiaojuan ZHU
;
Bin WU
;
Tao WU
;
Kangchen ZHAO
;
Yiyue GE
;
Lunbiao CUI
Author Information
1. 江苏省疾病预防控制中心 国家卫生健康委员会肠道病原微生物重点实验室 江苏省新发突发重大传染病病原微生物重点实验室，南京 210009
- Keywords:
2019-nCoV;
NSP1;
Variant;
Bioinformatics analysis
- From:
Chinese Journal of Experimental and Clinical Virology
2023;37(6):600-605
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore the effect of significant variation in non-structural protein 1 (NSP1) of 2019 novel coronavirus (2019-nCoV) on binding to 5′UTR, and to provide clues for the development of antiviral drugs and vaccines.Methods:The bioinformatics analysis of 2019-nCoV genome database was conducted to select the amino acid variation sites (T12A, R124L, N128I, K141A, GHVMV82-86DEL and KSF141-143DEL) that may affect the structure of NSP1 and the ability binding to 5′UTR. PSIPRED online tool was used to analyze the secondary structure of the variants, mCSM-NA was used to predict their binding ability to RNA, and DynaMut webserver was used to analyze the influence of variants on protein stability. The variant plasmids were constructed and transfected into HEK-293T cells. The dual luciferase reporter gene assay and RNA binding protein immunoprecipitation (RIP) assay were used to detect the binding ability of the NSP1 variant for viral 5′UTR.Results:Bioinformatics analysis predicted that except for R124L mutation, the other five variants could change the secondary structure of protein, and the mutations of T12A, R124L, N128I and K141A could reduce the binding ability of RNA, while the mutations of T12A, R124L and N128I reduced the stability of protein. The experimental results showed that R124L, N128I, GHVMV82-86DEL and KSF141-143DEL significantly weakened the binding ability of NSP1 to 5′UTR.Conclusions:Some mutations or deletion of NSP1 amino acids could change the secondary structure and significantly weaken the binding ability of NSP1 to the 5′UTR, suggesting that the pathogenicity of the virus may be changed, which could provide a theoretical basis for the development of antiviral drugs and vaccines.
