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Substitutions at Loop Regions of TMUV E Protein Domain III Differentially Impair Viral Entry and Assembly
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...
Autores principales: | , , , , , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8273266/ https://www.ncbi.nlm.nih.gov/pubmed/34262547 http://dx.doi.org/10.3389/fmicb.2021.688172 |
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author | Hu, Tao Wu, Zhen Wu, Shaoxiong Wang, Mingshu Jia, Renyong Zhu, Dekang Liu, Mafeng Zhao, Xinxin Yang, Qiao Wu, Ying Zhang, Shaqiu Huang, Juan Mao, Sai Ou, Xumin Gao, Qun Sun, Di Liu, Yunya Zhang, Ling Yu, YanLing Chen, Shun Cheng, Anchun |
author_facet | Hu, Tao Wu, Zhen Wu, Shaoxiong Wang, Mingshu Jia, Renyong Zhu, Dekang Liu, Mafeng Zhao, Xinxin Yang, Qiao Wu, Ying Zhang, Shaqiu Huang, Juan Mao, Sai Ou, Xumin Gao, Qun Sun, Di Liu, Yunya Zhang, Ling Yu, YanLing Chen, Shun Cheng, Anchun |
author_sort | Hu, Tao |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-8273266 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-82732662021-07-13 Substitutions at Loop Regions of TMUV E Protein Domain III Differentially Impair Viral Entry and Assembly Hu, Tao Wu, Zhen Wu, Shaoxiong Wang, Mingshu Jia, Renyong Zhu, Dekang Liu, Mafeng Zhao, Xinxin Yang, Qiao Wu, Ying Zhang, Shaqiu Huang, Juan Mao, Sai Ou, Xumin Gao, Qun Sun, Di Liu, Yunya Zhang, Ling Yu, YanLing Chen, Shun Cheng, Anchun Front Microbiol Microbiology 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. Frontiers Media S.A. 2021-06-28 /pmc/articles/PMC8273266/ /pubmed/34262547 http://dx.doi.org/10.3389/fmicb.2021.688172 Text en Copyright © 2021 Hu, Wu, Wu, Wang, Jia, Zhu, Liu, Zhao, Yang, Wu, Zhang, Huang, Mao, Ou, Gao, Sun, Liu, Zhang, Yu, Chen and Cheng. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Microbiology Hu, Tao Wu, Zhen Wu, Shaoxiong Wang, Mingshu Jia, Renyong Zhu, Dekang Liu, Mafeng Zhao, Xinxin Yang, Qiao Wu, Ying Zhang, Shaqiu Huang, Juan Mao, Sai Ou, Xumin Gao, Qun Sun, Di Liu, Yunya Zhang, Ling Yu, YanLing Chen, Shun Cheng, Anchun Substitutions at Loop Regions of TMUV E Protein Domain III Differentially Impair Viral Entry and Assembly |
title | Substitutions at Loop Regions of TMUV E Protein Domain III Differentially Impair Viral Entry and Assembly |
title_full | Substitutions at Loop Regions of TMUV E Protein Domain III Differentially Impair Viral Entry and Assembly |
title_fullStr | Substitutions at Loop Regions of TMUV E Protein Domain III Differentially Impair Viral Entry and Assembly |
title_full_unstemmed | Substitutions at Loop Regions of TMUV E Protein Domain III Differentially Impair Viral Entry and Assembly |
title_short | Substitutions at Loop Regions of TMUV E Protein Domain III Differentially Impair Viral Entry and Assembly |
title_sort | substitutions at loop regions of tmuv e protein domain iii differentially impair viral entry and assembly |
topic | Microbiology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8273266/ https://www.ncbi.nlm.nih.gov/pubmed/34262547 http://dx.doi.org/10.3389/fmicb.2021.688172 |
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