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Determinants of duck Tembusu virus NS2A/2B polyprotein procession attenuated viral replication and proliferation in vitro

Duck Tembusu virus (DTMUV), a mosquito-borne Flavivirus, has caused serious economic losses for the Chinese poultry industry. The genome is translated into a polyprotein that is cleaved to mature protein by host and viral proteases in the host cell, and this proteolytic process is important for the...

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Autores principales: Jiang, Bowen, Zhang, Wei, Wu, Yuanyuan, Wang, Tao, Wang, Mingshu, Jia, Renyong, Zhu, Dekang, Liu, Mafeng, Zhao, Xinxin, Yang, Qiao, Wu, Ying, Zhang, ShaQiu, Liu, YunYa, Zhang, Ling, Yu, YanLing, Pan, Leichang, Chen, Shun, Cheng, Anchun
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7381675/
https://www.ncbi.nlm.nih.gov/pubmed/32709930
http://dx.doi.org/10.1038/s41598-020-68271-0
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author Jiang, Bowen
Zhang, Wei
Wu, Yuanyuan
Wang, Tao
Wang, Mingshu
Jia, Renyong
Zhu, Dekang
Liu, Mafeng
Zhao, Xinxin
Yang, Qiao
Wu, Ying
Zhang, ShaQiu
Liu, YunYa
Zhang, Ling
Yu, YanLing
Pan, Leichang
Chen, Shun
Cheng, Anchun
author_facet Jiang, Bowen
Zhang, Wei
Wu, Yuanyuan
Wang, Tao
Wang, Mingshu
Jia, Renyong
Zhu, Dekang
Liu, Mafeng
Zhao, Xinxin
Yang, Qiao
Wu, Ying
Zhang, ShaQiu
Liu, YunYa
Zhang, Ling
Yu, YanLing
Pan, Leichang
Chen, Shun
Cheng, Anchun
author_sort Jiang, Bowen
collection PubMed
description Duck Tembusu virus (DTMUV), a mosquito-borne Flavivirus, has caused serious economic losses for the Chinese poultry industry. The genome is translated into a polyprotein that is cleaved to mature protein by host and viral proteases in the host cell, and this proteolytic process is important for the viral life cycle. However, the cleavage mechanism of DTMUV polyprotein is still unclear. In this study, we identified that several amino acids (P1-R, P1′-G, P2-R, P3-T, and P4-V) were vital for NS2A/2B cleavage. Meanwhile, both NS2A and NS2B were essential in cis for polyprotein NS2A/2B intramolecular cleavage. Subsequently, a DTMUV replicon and an infectious clone showed that the P1 site is essential to viral replication, while a mutation in P1′ could boost viral RNA replication. Furthermore, a recombinant virus with P1 and P1′ site mutations named rDTMUV-NS2A/2B-P1P1′(AA) was rescued from transfected BHK21 cells. The maximum viral titers and viral genome copies of rDTMUV-NS2A/2B-P1P1′(AA) were much lower than those of rDTMUV-WT both in the intracellular and extracellular samples of transfected and infected BHK21 cells. Taken together, the NS2A/2B cleavage sites processed by the NS2B3 protease are vital for DTMUV proliferation and virulence.
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spelling pubmed-73816752020-07-28 Determinants of duck Tembusu virus NS2A/2B polyprotein procession attenuated viral replication and proliferation in vitro Jiang, Bowen Zhang, Wei Wu, Yuanyuan Wang, Tao Wang, Mingshu Jia, Renyong Zhu, Dekang Liu, Mafeng Zhao, Xinxin Yang, Qiao Wu, Ying Zhang, ShaQiu Liu, YunYa Zhang, Ling Yu, YanLing Pan, Leichang Chen, Shun Cheng, Anchun Sci Rep Article Duck Tembusu virus (DTMUV), a mosquito-borne Flavivirus, has caused serious economic losses for the Chinese poultry industry. The genome is translated into a polyprotein that is cleaved to mature protein by host and viral proteases in the host cell, and this proteolytic process is important for the viral life cycle. However, the cleavage mechanism of DTMUV polyprotein is still unclear. In this study, we identified that several amino acids (P1-R, P1′-G, P2-R, P3-T, and P4-V) were vital for NS2A/2B cleavage. Meanwhile, both NS2A and NS2B were essential in cis for polyprotein NS2A/2B intramolecular cleavage. Subsequently, a DTMUV replicon and an infectious clone showed that the P1 site is essential to viral replication, while a mutation in P1′ could boost viral RNA replication. Furthermore, a recombinant virus with P1 and P1′ site mutations named rDTMUV-NS2A/2B-P1P1′(AA) was rescued from transfected BHK21 cells. The maximum viral titers and viral genome copies of rDTMUV-NS2A/2B-P1P1′(AA) were much lower than those of rDTMUV-WT both in the intracellular and extracellular samples of transfected and infected BHK21 cells. Taken together, the NS2A/2B cleavage sites processed by the NS2B3 protease are vital for DTMUV proliferation and virulence. Nature Publishing Group UK 2020-07-24 /pmc/articles/PMC7381675/ /pubmed/32709930 http://dx.doi.org/10.1038/s41598-020-68271-0 Text en © The Author(s) 2020, corrected publication 2021 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Jiang, Bowen
Zhang, Wei
Wu, Yuanyuan
Wang, Tao
Wang, Mingshu
Jia, Renyong
Zhu, Dekang
Liu, Mafeng
Zhao, Xinxin
Yang, Qiao
Wu, Ying
Zhang, ShaQiu
Liu, YunYa
Zhang, Ling
Yu, YanLing
Pan, Leichang
Chen, Shun
Cheng, Anchun
Determinants of duck Tembusu virus NS2A/2B polyprotein procession attenuated viral replication and proliferation in vitro
title Determinants of duck Tembusu virus NS2A/2B polyprotein procession attenuated viral replication and proliferation in vitro
title_full Determinants of duck Tembusu virus NS2A/2B polyprotein procession attenuated viral replication and proliferation in vitro
title_fullStr Determinants of duck Tembusu virus NS2A/2B polyprotein procession attenuated viral replication and proliferation in vitro
title_full_unstemmed Determinants of duck Tembusu virus NS2A/2B polyprotein procession attenuated viral replication and proliferation in vitro
title_short Determinants of duck Tembusu virus NS2A/2B polyprotein procession attenuated viral replication and proliferation in vitro
title_sort determinants of duck tembusu virus ns2a/2b polyprotein procession attenuated viral replication and proliferation in vitro
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7381675/
https://www.ncbi.nlm.nih.gov/pubmed/32709930
http://dx.doi.org/10.1038/s41598-020-68271-0
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