Cargando…

The Nsp12-coding region of type 2 PRRSV is required for viral subgenomic mRNA synthesis

As one of many nonstructural proteins of porcine reproductive and respiratory syndrome virus (PRRSV), nonstructural protein 12 (Nsp12) has received relatively little attention, and its role in virus replication, if any, is essentially unknown. By the application of reverse genetic manipulation of an...

Descripción completa

Detalles Bibliográficos
Autores principales: Wang, Tong-Yun, Fang, Qiong-Qiong, Cong, Feng, Liu, Yong-Gang, Wang, Hai-Ming, Zhang, Hong-Liang, Tian, Zhi-Jun, Tang, Yan-Dong, Cai, Xue-Hui
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Taylor & Francis 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6818116/
https://www.ncbi.nlm.nih.gov/pubmed/31631782
http://dx.doi.org/10.1080/22221751.2019.1679010
_version_ 1783463560498642944
author Wang, Tong-Yun
Fang, Qiong-Qiong
Cong, Feng
Liu, Yong-Gang
Wang, Hai-Ming
Zhang, Hong-Liang
Tian, Zhi-Jun
Tang, Yan-Dong
Cai, Xue-Hui
author_facet Wang, Tong-Yun
Fang, Qiong-Qiong
Cong, Feng
Liu, Yong-Gang
Wang, Hai-Ming
Zhang, Hong-Liang
Tian, Zhi-Jun
Tang, Yan-Dong
Cai, Xue-Hui
author_sort Wang, Tong-Yun
collection PubMed
description As one of many nonstructural proteins of porcine reproductive and respiratory syndrome virus (PRRSV), nonstructural protein 12 (Nsp12) has received relatively little attention, and its role in virus replication, if any, is essentially unknown. By the application of reverse genetic manipulation of an infectious PRRSV clone, the current study is the first to demonstrate that Nsp12 is a key component of PRRSV replication. In addition, the biochemical properties of Nsp12 were evaluated, revealing that Nsp12 forms dimers when exposed to oxidative conditions. Furthermore, we systemically analyzed the function of Nsp12 in PRRSV RNA synthesis using a strand-specific PCR method. To our surprise, Nsp12 was not found to be involved in minus-strand genomic RNA (-gRNA) synthesis; importantly, our results indicate that Nsp12 is involved in the synthesis of both plus- and minus-strand subgenomic mRNAs (+sgmRNA and -sgmRNA). Finally, we found that the combination of cysteine 35 and cysteine 79 in Nsp12 is required for sgmRNA synthesis. To our knowledge, we are the first to report the biological role of Nsp12 in the PRRSV lifecycle, and we conclude that Nsp12 is involved in the synthesis of both + sgRNA and -sgRNA.
format Online
Article
Text
id pubmed-6818116
institution National Center for Biotechnology Information
language English
publishDate 2019
publisher Taylor & Francis
record_format MEDLINE/PubMed
spelling pubmed-68181162019-11-05 The Nsp12-coding region of type 2 PRRSV is required for viral subgenomic mRNA synthesis Wang, Tong-Yun Fang, Qiong-Qiong Cong, Feng Liu, Yong-Gang Wang, Hai-Ming Zhang, Hong-Liang Tian, Zhi-Jun Tang, Yan-Dong Cai, Xue-Hui Emerg Microbes Infect Articles As one of many nonstructural proteins of porcine reproductive and respiratory syndrome virus (PRRSV), nonstructural protein 12 (Nsp12) has received relatively little attention, and its role in virus replication, if any, is essentially unknown. By the application of reverse genetic manipulation of an infectious PRRSV clone, the current study is the first to demonstrate that Nsp12 is a key component of PRRSV replication. In addition, the biochemical properties of Nsp12 were evaluated, revealing that Nsp12 forms dimers when exposed to oxidative conditions. Furthermore, we systemically analyzed the function of Nsp12 in PRRSV RNA synthesis using a strand-specific PCR method. To our surprise, Nsp12 was not found to be involved in minus-strand genomic RNA (-gRNA) synthesis; importantly, our results indicate that Nsp12 is involved in the synthesis of both plus- and minus-strand subgenomic mRNAs (+sgmRNA and -sgmRNA). Finally, we found that the combination of cysteine 35 and cysteine 79 in Nsp12 is required for sgmRNA synthesis. To our knowledge, we are the first to report the biological role of Nsp12 in the PRRSV lifecycle, and we conclude that Nsp12 is involved in the synthesis of both + sgRNA and -sgRNA. Taylor & Francis 2019-10-21 /pmc/articles/PMC6818116/ /pubmed/31631782 http://dx.doi.org/10.1080/22221751.2019.1679010 Text en © 2019 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group, on behalf of Shanghai Shangyixun Cultural Communication Co., Ltd https://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/ (https://creativecommons.org/licenses/by/4.0/) ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Articles
Wang, Tong-Yun
Fang, Qiong-Qiong
Cong, Feng
Liu, Yong-Gang
Wang, Hai-Ming
Zhang, Hong-Liang
Tian, Zhi-Jun
Tang, Yan-Dong
Cai, Xue-Hui
The Nsp12-coding region of type 2 PRRSV is required for viral subgenomic mRNA synthesis
title The Nsp12-coding region of type 2 PRRSV is required for viral subgenomic mRNA synthesis
title_full The Nsp12-coding region of type 2 PRRSV is required for viral subgenomic mRNA synthesis
title_fullStr The Nsp12-coding region of type 2 PRRSV is required for viral subgenomic mRNA synthesis
title_full_unstemmed The Nsp12-coding region of type 2 PRRSV is required for viral subgenomic mRNA synthesis
title_short The Nsp12-coding region of type 2 PRRSV is required for viral subgenomic mRNA synthesis
title_sort nsp12-coding region of type 2 prrsv is required for viral subgenomic mrna synthesis
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6818116/
https://www.ncbi.nlm.nih.gov/pubmed/31631782
http://dx.doi.org/10.1080/22221751.2019.1679010
work_keys_str_mv AT wangtongyun thensp12codingregionoftype2prrsvisrequiredforviralsubgenomicmrnasynthesis
AT fangqiongqiong thensp12codingregionoftype2prrsvisrequiredforviralsubgenomicmrnasynthesis
AT congfeng thensp12codingregionoftype2prrsvisrequiredforviralsubgenomicmrnasynthesis
AT liuyonggang thensp12codingregionoftype2prrsvisrequiredforviralsubgenomicmrnasynthesis
AT wanghaiming thensp12codingregionoftype2prrsvisrequiredforviralsubgenomicmrnasynthesis
AT zhanghongliang thensp12codingregionoftype2prrsvisrequiredforviralsubgenomicmrnasynthesis
AT tianzhijun thensp12codingregionoftype2prrsvisrequiredforviralsubgenomicmrnasynthesis
AT tangyandong thensp12codingregionoftype2prrsvisrequiredforviralsubgenomicmrnasynthesis
AT caixuehui thensp12codingregionoftype2prrsvisrequiredforviralsubgenomicmrnasynthesis
AT wangtongyun nsp12codingregionoftype2prrsvisrequiredforviralsubgenomicmrnasynthesis
AT fangqiongqiong nsp12codingregionoftype2prrsvisrequiredforviralsubgenomicmrnasynthesis
AT congfeng nsp12codingregionoftype2prrsvisrequiredforviralsubgenomicmrnasynthesis
AT liuyonggang nsp12codingregionoftype2prrsvisrequiredforviralsubgenomicmrnasynthesis
AT wanghaiming nsp12codingregionoftype2prrsvisrequiredforviralsubgenomicmrnasynthesis
AT zhanghongliang nsp12codingregionoftype2prrsvisrequiredforviralsubgenomicmrnasynthesis
AT tianzhijun nsp12codingregionoftype2prrsvisrequiredforviralsubgenomicmrnasynthesis
AT tangyandong nsp12codingregionoftype2prrsvisrequiredforviralsubgenomicmrnasynthesis
AT caixuehui nsp12codingregionoftype2prrsvisrequiredforviralsubgenomicmrnasynthesis