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Zika virus NS3 is a canonical RNA helicase stimulated by NS5 RNA polymerase

Zika virus is a positive single-strand RNA virus whose replication involved RNA unwinding and synthesis. ZIKV NS3 contains a helicase domain, but its enzymatic activity is not fully characterized. Here, we established a dsRNA unwinding assay based on the FRET effect to study the helicase activity of...

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Detalles Bibliográficos
Autores principales: Xu, Shan, Ci, Yali, Wang, Leijie, Yang, Yang, Zhang, Leiliang, Xu, Caimin, Qin, Chengfeng, Shi, Lei
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6895266/
https://www.ncbi.nlm.nih.gov/pubmed/31361901
http://dx.doi.org/10.1093/nar/gkz650
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author Xu, Shan
Ci, Yali
Wang, Leijie
Yang, Yang
Zhang, Leiliang
Xu, Caimin
Qin, Chengfeng
Shi, Lei
author_facet Xu, Shan
Ci, Yali
Wang, Leijie
Yang, Yang
Zhang, Leiliang
Xu, Caimin
Qin, Chengfeng
Shi, Lei
author_sort Xu, Shan
collection PubMed
description Zika virus is a positive single-strand RNA virus whose replication involved RNA unwinding and synthesis. ZIKV NS3 contains a helicase domain, but its enzymatic activity is not fully characterized. Here, we established a dsRNA unwinding assay based on the FRET effect to study the helicase activity of ZIKV NS3, which provided kinetic information in real time. We found that ZIKV NS3 specifically unwound dsRNA/dsDNA with a 3′ overhang in the 3′ to 5′ direction. The RNA unwinding ability of NS3 significantly decreased when the duplex was longer than 18 base pairs. The helicase activity of NS3 depends on ATP hydrolysis and binding to RNA. Mutations in the ATP binding region or the RNA binding region of NS3 impair its helicase activity, thus blocking viral replication in the cell. Furthermore, we showed that ZIKV NS5 interacted with NS3 and stimulated its helicase activity. Disrupting NS3-NS5 interaction resulted in a defect in viral replication, revealing the tight coupling of RNA unwinding and synthesis. We suggest that NS3 helicase activity is stimulated by NS5; thus, viral replication can be carried out efficiently. Our work provides a molecular mechanism of ZIKV NS3 unwinding and novel insights into ZIKV replication.
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spelling pubmed-68952662019-12-11 Zika virus NS3 is a canonical RNA helicase stimulated by NS5 RNA polymerase Xu, Shan Ci, Yali Wang, Leijie Yang, Yang Zhang, Leiliang Xu, Caimin Qin, Chengfeng Shi, Lei Nucleic Acids Res Nucleic Acid Enzymes Zika virus is a positive single-strand RNA virus whose replication involved RNA unwinding and synthesis. ZIKV NS3 contains a helicase domain, but its enzymatic activity is not fully characterized. Here, we established a dsRNA unwinding assay based on the FRET effect to study the helicase activity of ZIKV NS3, which provided kinetic information in real time. We found that ZIKV NS3 specifically unwound dsRNA/dsDNA with a 3′ overhang in the 3′ to 5′ direction. The RNA unwinding ability of NS3 significantly decreased when the duplex was longer than 18 base pairs. The helicase activity of NS3 depends on ATP hydrolysis and binding to RNA. Mutations in the ATP binding region or the RNA binding region of NS3 impair its helicase activity, thus blocking viral replication in the cell. Furthermore, we showed that ZIKV NS5 interacted with NS3 and stimulated its helicase activity. Disrupting NS3-NS5 interaction resulted in a defect in viral replication, revealing the tight coupling of RNA unwinding and synthesis. We suggest that NS3 helicase activity is stimulated by NS5; thus, viral replication can be carried out efficiently. Our work provides a molecular mechanism of ZIKV NS3 unwinding and novel insights into ZIKV replication. Oxford University Press 2019-09-19 2019-07-30 /pmc/articles/PMC6895266/ /pubmed/31361901 http://dx.doi.org/10.1093/nar/gkz650 Text en © The Author(s) 2019. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Nucleic Acid Enzymes
Xu, Shan
Ci, Yali
Wang, Leijie
Yang, Yang
Zhang, Leiliang
Xu, Caimin
Qin, Chengfeng
Shi, Lei
Zika virus NS3 is a canonical RNA helicase stimulated by NS5 RNA polymerase
title Zika virus NS3 is a canonical RNA helicase stimulated by NS5 RNA polymerase
title_full Zika virus NS3 is a canonical RNA helicase stimulated by NS5 RNA polymerase
title_fullStr Zika virus NS3 is a canonical RNA helicase stimulated by NS5 RNA polymerase
title_full_unstemmed Zika virus NS3 is a canonical RNA helicase stimulated by NS5 RNA polymerase
title_short Zika virus NS3 is a canonical RNA helicase stimulated by NS5 RNA polymerase
title_sort zika virus ns3 is a canonical rna helicase stimulated by ns5 rna polymerase
topic Nucleic Acid Enzymes
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6895266/
https://www.ncbi.nlm.nih.gov/pubmed/31361901
http://dx.doi.org/10.1093/nar/gkz650
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