Effect of mutations in the key amino acids of enterovirus 71 3D protein on virus replication
10.3760/cma.j.cn112866-20210622-00103
- VernacularTitle:肠道病毒71型3D蛋白关键氨基酸突变对病毒增殖的影响
- Author:
Xiaoying XU
1
;
Hailu ZHANG
;
Yulei ZHANG
;
Yao WANG
;
Xuemin WEI
;
Haowen YUAN
;
Hao LIANG
;
Li ZHAO
;
Zhiyu WANG
;
Hongling WEN
Author Information
1. 山东大学齐鲁医学院公共卫生学院微生物检验学系 山东省"十三五"高等学校感染性疾病防控重点实验室，济南 250012
- Keywords:
3D protein of enterovirus 71;
Mutations;
Replication;
The tertiary structure
- From:
Chinese Journal of Experimental and Clinical Virology
2022;36(1):29-33
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore the effect of key amino acid mutations of 3D protein in enterovirus 71 on viral proliferation, and infer the mechanism of mutation affecting viral proliferation according to the tertiary structure model of 3D protein.Methods:The proliferation characteristics of the mutant strain that was constructed by site-directed mutagenesis and reverse genetics using the pMD19T-SDLY107-EGFP constructed from the fatal strain SDLY107 as a template were measured. Meanwhile the tertiary structure of the 3D protein was predicted, and then the possible mechanism of the mutation affecting the proliferation ability of the virus was speculated according to the functional characteristics of the domain of the 3D protein.Results:Two mutant viruses, eGFP-EV-A71 (S37N) and eGFP-EV-A71 (R142K), were successfully constructed by double enzyme digestion and viral gene sequencing. The proliferation rate of the two mutant strains in RD cells was significantly lower than that of the parent strains. The 3D protein tertiary structure prediction model showed that the 3D protein consisted of three domains: "Finger" , "thumb" and "Palm" , constituting a cupped right-handed structure. S37N and R142K are located in the "thumb" domain and " finger" domain, respectively. The "thumb" and "finger" domains have important effects on the activity of 3D polymerase and the stability of protein.Conclusions:Mutations at S37N and R142K sites of EV71 3D protein decrease the replication ability of EV71, and these two mutations may affect the proliferation of EV71 virus by changing the 3D protein polymerase activity and the interactions between multiple domains.
