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...
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| Veröffentlicht in: | Emerging microbes & infections 2019-01, Vol.8 (1), p.1501-1510 |
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| creator | 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 |
| 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. |
| doi_str_mv | 10.1080/22221751.2019.1679010 |
| format | Article |
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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.</description><identifier>ISSN: 2222-1751</identifier><identifier>EISSN: 2222-1751</identifier><identifier>DOI: 10.1080/22221751.2019.1679010</identifier><identifier>PMID: 31631782</identifier><language>eng</language><publisher>United States: Taylor & Francis</publisher><subject>Animals ; dimer ; Gene Expression Regulation, Viral ; Nsp12 ; Open Reading Frames ; Porcine Reproductive and Respiratory Syndrome - virology ; Porcine respiratory and reproductive syndrome virus - genetics ; Porcine respiratory and reproductive syndrome virus - metabolism ; PRRSV ; RNA synthesis ; RNA, Messenger - genetics ; RNA, Messenger - metabolism ; RNA, Viral - genetics ; RNA, Viral - metabolism ; subgenomic mRNA ; Swine ; Transcription, Genetic ; Viral Nonstructural Proteins - genetics ; Viral Nonstructural Proteins - metabolism ; Virus Replication</subject><ispartof>Emerging microbes & infections, 2019-01, Vol.8 (1), p.1501-1510</ispartof><rights>2019 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group, on behalf of Shanghai Shangyixun Cultural Communication Co., Ltd 2019</rights><rights>2019 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group, on behalf of Shanghai Shangyixun Cultural Communication Co., Ltd. This work is licensed under the Creative Commons Attribution License http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2019 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group, on behalf of Shanghai Shangyixun Cultural Communication Co., Ltd 2019 The Author(s)</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c562t-28f374cc25dd489147806133fba2c86912fc0774b4e9c6a894ab03697ca6a6d33</citedby><cites>FETCH-LOGICAL-c562t-28f374cc25dd489147806133fba2c86912fc0774b4e9c6a894ab03697ca6a6d33</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC6818116/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC6818116/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,864,885,2102,27502,27924,27925,53791,53793,59143,59144</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/31631782$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Wang, Tong-Yun</creatorcontrib><creatorcontrib>Fang, Qiong-Qiong</creatorcontrib><creatorcontrib>Cong, Feng</creatorcontrib><creatorcontrib>Liu, Yong-Gang</creatorcontrib><creatorcontrib>Wang, Hai-Ming</creatorcontrib><creatorcontrib>Zhang, Hong-Liang</creatorcontrib><creatorcontrib>Tian, Zhi-Jun</creatorcontrib><creatorcontrib>Tang, Yan-Dong</creatorcontrib><creatorcontrib>Cai, Xue-Hui</creatorcontrib><title>The Nsp12-coding region of type 2 PRRSV is required for viral subgenomic mRNA synthesis</title><title>Emerging microbes & infections</title><addtitle>Emerg Microbes Infect</addtitle><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.</description><subject>Animals</subject><subject>dimer</subject><subject>Gene Expression Regulation, Viral</subject><subject>Nsp12</subject><subject>Open Reading Frames</subject><subject>Porcine Reproductive and Respiratory Syndrome - virology</subject><subject>Porcine respiratory and reproductive syndrome virus - genetics</subject><subject>Porcine respiratory and reproductive syndrome virus - metabolism</subject><subject>PRRSV</subject><subject>RNA synthesis</subject><subject>RNA, Messenger - genetics</subject><subject>RNA, Messenger - metabolism</subject><subject>RNA, Viral - genetics</subject><subject>RNA, Viral - metabolism</subject><subject>subgenomic mRNA</subject><subject>Swine</subject><subject>Transcription, Genetic</subject><subject>Viral Nonstructural Proteins - genetics</subject><subject>Viral Nonstructural Proteins - metabolism</subject><subject>Virus Replication</subject><issn>2222-1751</issn><issn>2222-1751</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2019</creationdate><recordtype>article</recordtype><sourceid>0YH</sourceid><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>DOA</sourceid><recordid>eNp9Uk1vEzEQtRCIVqE_AWSJC5cN_lrbe0FUVYFKValCgaPl9dqJo107tXeL8u9xSFq1HDoXW_PePHtmHgBvMZpjJNFHUgKLGs8Jws0cc9EgjF6A412-2gEvH92PwEnOa1RCIM4wew2OKOYUC0mOwe-blYVXeYNJZWLnwxImu_QxwOjguN1YSOD1YvHjF_S5ILeTT7aDLiZ455PuYZ7apQ1x8AYOi6tTmLdhXNns8xvwyuk-25PDOQM_v5zfnH2rLr9_vTg7vaxMzclYEemoYMaQuuuYbDATEnFMqWs1MZI3mDiDhGAts43hWjZMt4jyRhjNNe8onYGLvW4X9Vptkh902qqovfqXiGmpdBq96a0yjDHetZgQ5hghSDqJ60ZqYY2Umrii9WmvtZnawXbGhrH0-ET0KRL8Si3jneISS1xGOgMfDgIp3k42j2rw2di-18HGKStCkaAN5TUu1Pf_UddxSqGMShFGRE1qwUVh1XuWSTHnZN3DZzBSOyeoeyeonRPUwQml7t3jTh6q7vdeCJ_3BB_KMgf9J6a-U6Pe9jG5pIPxuZCffeMvTcm_ig</recordid><startdate>20190101</startdate><enddate>20190101</enddate><creator>Wang, Tong-Yun</creator><creator>Fang, Qiong-Qiong</creator><creator>Cong, Feng</creator><creator>Liu, Yong-Gang</creator><creator>Wang, Hai-Ming</creator><creator>Zhang, Hong-Liang</creator><creator>Tian, Zhi-Jun</creator><creator>Tang, Yan-Dong</creator><creator>Cai, Xue-Hui</creator><general>Taylor & Francis</general><general>Taylor & Francis Ltd</general><general>Taylor & Francis Group</general><scope>0YH</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BENPR</scope><scope>CCPQU</scope><scope>COVID</scope><scope>DWQXO</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>K9.</scope><scope>M0S</scope><scope>M1P</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>7X8</scope><scope>5PM</scope><scope>DOA</scope></search><sort><creationdate>20190101</creationdate><title>The Nsp12-coding region of type 2 PRRSV is required for viral subgenomic mRNA synthesis</title><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></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c562t-28f374cc25dd489147806133fba2c86912fc0774b4e9c6a894ab03697ca6a6d33</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2019</creationdate><topic>Animals</topic><topic>dimer</topic><topic>Gene Expression Regulation, Viral</topic><topic>Nsp12</topic><topic>Open Reading Frames</topic><topic>Porcine Reproductive and Respiratory Syndrome - virology</topic><topic>Porcine respiratory and reproductive syndrome virus - genetics</topic><topic>Porcine respiratory and reproductive syndrome virus - metabolism</topic><topic>PRRSV</topic><topic>RNA synthesis</topic><topic>RNA, Messenger - genetics</topic><topic>RNA, Messenger - metabolism</topic><topic>RNA, Viral - genetics</topic><topic>RNA, Viral - metabolism</topic><topic>subgenomic mRNA</topic><topic>Swine</topic><topic>Transcription, Genetic</topic><topic>Viral Nonstructural Proteins - genetics</topic><topic>Viral Nonstructural Proteins - metabolism</topic><topic>Virus Replication</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Wang, Tong-Yun</creatorcontrib><creatorcontrib>Fang, Qiong-Qiong</creatorcontrib><creatorcontrib>Cong, Feng</creatorcontrib><creatorcontrib>Liu, Yong-Gang</creatorcontrib><creatorcontrib>Wang, Hai-Ming</creatorcontrib><creatorcontrib>Zhang, Hong-Liang</creatorcontrib><creatorcontrib>Tian, Zhi-Jun</creatorcontrib><creatorcontrib>Tang, Yan-Dong</creatorcontrib><creatorcontrib>Cai, Xue-Hui</creatorcontrib><collection>Taylor & Francis (Open access)</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Health Medical collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni)</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central Essentials</collection><collection>ProQuest Central</collection><collection>ProQuest One Community College</collection><collection>Coronavirus Research Database</collection><collection>ProQuest Central</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Publicly Available Content Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>Emerging microbes & infections</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Wang, Tong-Yun</au><au>Fang, Qiong-Qiong</au><au>Cong, Feng</au><au>Liu, Yong-Gang</au><au>Wang, Hai-Ming</au><au>Zhang, Hong-Liang</au><au>Tian, Zhi-Jun</au><au>Tang, Yan-Dong</au><au>Cai, Xue-Hui</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>The Nsp12-coding region of type 2 PRRSV is required for viral subgenomic mRNA synthesis</atitle><jtitle>Emerging microbes & infections</jtitle><addtitle>Emerg Microbes Infect</addtitle><date>2019-01-01</date><risdate>2019</risdate><volume>8</volume><issue>1</issue><spage>1501</spage><epage>1510</epage><pages>1501-1510</pages><issn>2222-1751</issn><eissn>2222-1751</eissn><abstract>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.</abstract><cop>United States</cop><pub>Taylor & Francis</pub><pmid>31631782</pmid><doi>10.1080/22221751.2019.1679010</doi><tpages>10</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Animals dimer Gene Expression Regulation, Viral Nsp12 Open Reading Frames Porcine Reproductive and Respiratory Syndrome - virology Porcine respiratory and reproductive syndrome virus - genetics Porcine respiratory and reproductive syndrome virus - metabolism PRRSV RNA synthesis RNA, Messenger - genetics RNA, Messenger - metabolism RNA, Viral - genetics RNA, Viral - metabolism subgenomic mRNA Swine Transcription, Genetic Viral Nonstructural Proteins - genetics Viral Nonstructural Proteins - metabolism Virus Replication |
| title | The Nsp12-coding region of type 2 PRRSV is required for viral subgenomic mRNA synthesis |
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