Frontiers in Microbiology (Jun 2021)
Substitutions at Loop Regions of TMUV E Protein Domain III Differentially Impair Viral Entry and Assembly
- Tao Hu,
- Zhen Wu,
- Shaoxiong Wu,
- Mingshu Wang,
- Mingshu Wang,
- Mingshu Wang,
- Renyong Jia,
- Renyong Jia,
- Renyong Jia,
- Dekang Zhu,
- Dekang Zhu,
- Dekang Zhu,
- Mafeng Liu,
- Mafeng Liu,
- Mafeng Liu,
- Xinxin Zhao,
- Xinxin Zhao,
- Xinxin Zhao,
- Qiao Yang,
- Qiao Yang,
- Qiao Yang,
- Ying Wu,
- Ying Wu,
- Ying Wu,
- Shaqiu Zhang,
- Shaqiu Zhang,
- Shaqiu Zhang,
- Juan Huang,
- Juan Huang,
- Juan Huang,
- Sai Mao,
- Sai Mao,
- Sai Mao,
- Xumin Ou,
- Xumin Ou,
- Xumin Ou,
- Qun Gao,
- Qun Gao,
- Qun Gao,
- Di Sun,
- Di Sun,
- Di Sun,
- Yunya Liu,
- Ling Zhang,
- YanLing Yu,
- Shun Chen,
- Shun Chen,
- Shun Chen,
- Anchun Cheng,
- Anchun Cheng,
- Anchun Cheng
Affiliations
- Tao Hu
- Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, China
- Zhen Wu
- Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, China
- Shaoxiong Wu
- Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, China
- Mingshu Wang
- Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, China
- Mingshu Wang
- Institute of Preventive Veterinary Medicine, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, China
- Mingshu Wang
- Key Laboratory of Animal Disease and Human Health of Sichuan Province, Chengdu, China
- Renyong Jia
- Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, China
- Renyong Jia
- Institute of Preventive Veterinary Medicine, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, China
- Renyong Jia
- Key Laboratory of Animal Disease and Human Health of Sichuan Province, Chengdu, China
- Dekang Zhu
- Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, China
- Dekang Zhu
- Institute of Preventive Veterinary Medicine, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, China
- Dekang Zhu
- Key Laboratory of Animal Disease and Human Health of Sichuan Province, Chengdu, China
- Mafeng Liu
- Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, China
- Mafeng Liu
- Institute of Preventive Veterinary Medicine, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, China
- Mafeng Liu
- Key Laboratory of Animal Disease and Human Health of Sichuan Province, Chengdu, China
- Xinxin Zhao
- Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, China
- Xinxin Zhao
- Institute of Preventive Veterinary Medicine, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, China
- Xinxin Zhao
- Key Laboratory of Animal Disease and Human Health of Sichuan Province, Chengdu, China
- Qiao Yang
- Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, China
- Qiao Yang
- Institute of Preventive Veterinary Medicine, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, China
- Qiao Yang
- Key Laboratory of Animal Disease and Human Health of Sichuan Province, Chengdu, China
- Ying Wu
- Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, China
- Ying Wu
- Institute of Preventive Veterinary Medicine, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, China
- Ying Wu
- Key Laboratory of Animal Disease and Human Health of Sichuan Province, Chengdu, China
- Shaqiu Zhang
- Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, China
- Shaqiu Zhang
- Institute of Preventive Veterinary Medicine, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, China
- Shaqiu Zhang
- Key Laboratory of Animal Disease and Human Health of Sichuan Province, Chengdu, China
- Juan Huang
- Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, China
- Juan Huang
- Institute of Preventive Veterinary Medicine, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, China
- Juan Huang
- Key Laboratory of Animal Disease and Human Health of Sichuan Province, Chengdu, China
- Sai Mao
- Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, China
- Sai Mao
- Institute of Preventive Veterinary Medicine, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, China
- Sai Mao
- Key Laboratory of Animal Disease and Human Health of Sichuan Province, Chengdu, China
- Xumin Ou
- Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, China
- Xumin Ou
- Institute of Preventive Veterinary Medicine, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, China
- Xumin Ou
- Key Laboratory of Animal Disease and Human Health of Sichuan Province, Chengdu, China
- Qun Gao
- Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, China
- Qun Gao
- Institute of Preventive Veterinary Medicine, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, China
- Qun Gao
- Key Laboratory of Animal Disease and Human Health of Sichuan Province, Chengdu, China
- Di Sun
- Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, China
- Di Sun
- Institute of Preventive Veterinary Medicine, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, China
- Di Sun
- Key Laboratory of Animal Disease and Human Health of Sichuan Province, Chengdu, China
- Yunya Liu
- Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, China
- Ling Zhang
- Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, China
- YanLing Yu
- Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, China
- Shun Chen
- Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, China
- Shun Chen
- Institute of Preventive Veterinary Medicine, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, China
- Shun Chen
- Key Laboratory of Animal Disease and Human Health of Sichuan Province, Chengdu, China
- Anchun Cheng
- Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, China
- Anchun Cheng
- Institute of Preventive Veterinary Medicine, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, China
- Anchun Cheng
- Key Laboratory of Animal Disease and Human Health of Sichuan Province, Chengdu, China
- DOI
- https://doi.org/10.3389/fmicb.2021.688172
- Journal volume & issue
-
Vol. 12
Abstract
Flavivirus envelope protein (E) plays an important role in cellular infection, especially in virulence and antigenicity. E domain III of Tembusu virus (TMUV) is highly conserved among flaviviruses and contains four loop regions. However, the functions of the loop regions of TMUV E domain III in the viral life cycle have not yet been discovered. In this study, using a reverse genetics system, we performed site-directed mutagenesis on loops I, II, III, and IV of TMUV E domain III. Mutant 6 (S388A.G389A.K390A) showed better proliferation than the wild-type virus, while mutants 1–5 exhibited decreased in vitro infectivity, as determined by immunofluorescence assay (IFA). Based on a TMUV replicon system, the mutations exhibited no apparent effect on TMUV RNA replication. Subcellular fractionation assays and packaging system assays indicated that mutations in loops II–IV (T332A, T332S, S365A.S366A.T367A, and S388A.G389A.K390A, respectively) disrupted virion assembly. Moreover, loops I–IV played an important role in virus binding and entry, while mutant 6 (S388A.G389A.K390A) exhibited robust activity in virus entry. Taken together, our findings indicated the critical role of the loop regions in TMUV E domain III in the virus entry and assembly process.
Keywords