Nucleocytoplasmic shuttling of the West Nile virus        <fc >RNA</fc>RNA        ‐dependent        <fc >RNA</fc>RNA        polymerase        <fc >NS5</fc>NS5        is critical to infection

Lopez‐Denman Adam J.; Russo Alice; Wagstaff Kylie M.; White Peter A.; Jans David A.; Mackenzie Jason M.

摘要

Abstract > West Nile virus (WNV) is a single‐stranded, positive sense RNA virus of the family <fi>Flaviviridae</fi> and is a significant pathogen of global medical importance. Flavivirus replication is known to be exclusively cytoplasmic, but we show here for the first time that access to the nucleus of the WNV strain Kunjin (WNV <sub>KUN</sub> ) RNA‐dependent RNA polymerase (protein NS5) is central to WNV <sub>KUN</sub> virus production. We show that treatment of cells with the specific nuclear export inhibitor leptomycin B (LMB) results in increased NS5 nuclear accumulation in WNV <sub>KUN</sub> ‐infected cells and NS5‐transfected cells, indicative of nucleocytoplasmic shuttling under normal conditions. We used site‐directed mutagenesis to identify the nuclear localisation sequence (NLS) responsible for WNV <sub>KUN</sub> NS5 nuclear targeting, observing that mutation of this NLS resulted in exclusively cytoplasmic accumulation of NS5 even in the presence of leptomycin B. Introduction of NS5 NLS mutations into FLSDX, an infectious clone of WNV <sub>KUN</sub> , resulted in lethality, suggesting that the ability of NS5 to traffic into the nucleus in integral to WNV <sub>KUN</sub> replication. This study thus shows for the first time that NLS‐dependent trafficking into the nucleus during infection of WNV <sub>KUN</sub> NS5 is critical for viral replication. Excitingly, specific inhibitors of NS5 nuclear import reduce WNV <sub>KUN</sub> virus production, proving the principle that inhibition of WNV <sub>KUN</sub> NS5 nuclear import is a viable therapeutic avenue for antiviral drug development in the future. </abstract> </span> <span class="z_kbtn z_kbtnclass hoverxs" style="display: none;">展开▼</span> </div> <div class="translation abstracttxt"> <span class="zhankaihshouqi fivelineshidden" id="abstract"> <span>机译：</span><Abstract Type =“Main”XML：Lang =“en”> <标题类型=“main”>抽象</ title> >西尼罗病毒（Wnv）是一个单链，阳性感觉RNA病毒的家庭<fi> flaviviridae </ fi>是全球医学重要性的重要病原体。已知黄病毒复制是专门的细胞质，但我们首次在此处显示访问WNV菌株Kunjin的核（WnV <sub> kun </ sub>）RNA依赖性RNA聚合酶（蛋白质NS5）是中心的Wnv <sub> kun </ sub>病毒制作。我们表明，具有特定核导出抑制剂百霉素B（LMB）的细胞治疗导致WNV <Sub> kun </ sub>-infacted细胞和NS5转染细胞中的NS5核积累增加，指示正常条件下的核细胞骨质穿梭。我们使用定向诱变诱变核定位序列（NLS），其负责WNV <sub> kun </ sub> ns5核靶向，观察到这种NLS的突变即使在含有百霉素B的存在下，也导致NS5的特征细胞质积累。将NS5 NLS突变引入FLSDX中，WNV <sub> kun </ sub>的传染性克隆导致致死，表明NS5以Wnv <sub> kun的整体流入核的能力为kun </ sub>复制。因此，本研究表明，在WnV <sub> kun </ sub> ns5感染期间第一次表现出N1s依赖于核心的核，对于病毒复制至关重要。令人兴奋的是，NS5核导入的特异性抑制剂减少WNV <sub> kun </ sub>病毒的生产，证明了抑制Wnv <sub> kun </ sub> ns5核导入的原理是抗病毒药物开发的可行治疗途径未来。 </ p> </ abstract> </span> <span class="z_kbtn z_kbtnclass hoverxs" style="display: none;">展开▼</span> </div> </div> <div class="record"> <h2 class="all_title" id="enpatent33" >著录项</h2> <ul> <li> <span class="lefttit">来源</span> <div style="width: 86%;vertical-align: text-top;display: inline-block;"> <a href='/journal-foreign-15891/'>《Cellular microbiology》</a> <b style="margin: 0 2px;">|</b><span>2018年第8期</span><b style="margin: 0 2px;">|</b><span>共13页</span> </div> </li> <li> <div class="author"> <span class="lefttit">作者</span> <p id="fAuthorthree" class="threelineshidden zhankaihshouqi"> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Lopez‐Denman Adam J.&option=202" target="_blank" rel="nofollow">Lopez‐Denman Adam J.;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Russo Alice&option=202" target="_blank" rel="nofollow">Russo Alice;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Wagstaff Kylie M.&option=202" target="_blank" rel="nofollow">Wagstaff Kylie M.;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=White Peter A.&option=202" target="_blank" rel="nofollow">White Peter A.;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Jans David A.&option=202" target="_blank" rel="nofollow">Jans David A.;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Mackenzie Jason M.&option=202" target="_blank" rel="nofollow">Mackenzie Jason M.;</a> </p> <span class="z_kbtnclass z_kbtnclassall hoverxs" id="zkzz" style="display: none;">展开▼</span> </div> </li> <li> <div style="display: flex;"> <span class="lefttit">作者单位</span> <div style="position: relative;margin-left: 3px;max-width: 639px;"> <div class="threelineshidden zhankaihshouqi" id="fOrgthree"> <p>Department of Microbiology and Immunology Peter Doherty Institute for Infection and ImmunityUniversity of MelbourneMelbourne Victoria Australia;</p> <p>Faculty of Science Biotechnology and Biomolecular SciencesUniversity of New South WalesSydney New South Wales Australia;</p> <p>Department of BiochemistryMonash UniversityMelbourne Victoria Australia;</p> <p>Faculty of Science Biotechnology and Biomolecular SciencesUniversity of New South WalesSydney New South Wales Australia;</p> <p>Department of BiochemistryMonash UniversityMelbourne Victoria Australia;</p> <p>Department of Microbiology and Immunology Peter Doherty Institute for Infection and ImmunityUniversity of MelbourneMelbourne Victoria Australia;</p> </div> <span class="z_kbtnclass z_kbtnclassall hoverxs" id="zhdw" style="display: none;">展开▼</span> </div> </div> </li> <li > <span class="lefttit">收录信息</span> <span style="width: 86%;vertical-align: text-top;display: inline-block;"></span> </li> <li> <span class="lefttit">原文格式</span> <span>PDF</span> </li> <li> <span class="lefttit">正文语种</span> <span>eng</span> </li> <li> <span class="lefttit">中图分类</span> <span><a href="https://www.zhangqiaokeyan.com/clc/173.html" title="普通生物学">普通生物学;</a></span> </li> <li class="antistop"> <span class="lefttit">关键词</span> <p style="width: 86%;vertical-align: text-top;"> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=NS5&option=203" rel="nofollow">NS5;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=nuclear export&option=203" rel="nofollow">nuclear export;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=nuclear import&option=203" rel="nofollow">nuclear import;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=nucleus&option=203" rel="nofollow">nucleus;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=RNA ‐dependent RNA polymerase&option=203" rel="nofollow">RNA ‐dependent RNA polymerase;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=West Nile virus&option=203" rel="nofollow">West Nile virus;</a> </p> <div class="translation"> 机译：NS5;核导出;核导入;核;RNA依赖性RNA聚合酶;西尼罗河病毒; </div> </li> </ul> </div> </div> <div class="literature cardcommon"> <div class="similarity "> <h3 class="all_title" id="enpatent66">相似文献</h3> <div class="similaritytab clearfix"> <ul> <li class="active" >外文文献</li> <li >中文文献</li> <li >专利</li> </ul> </div> <div class="similarity_details"> <ul > <li> <div> <b>1. </b><a class="enjiyixqcontent" href="/journal-foreign-detail/0704022032057.html">Nucleocytoplasmic shuttling of the West Nile virus <fc >RNA</fc>RNA ‐dependent <fc >RNA</fc>RNA polymerase <fc >NS5</fc>NS5 is critical to infection</a> <b>[J]</b> . <span> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Lopez‐Denman Adam J.&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">Lopez‐Denman Adam 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Nucleocytoplasmic shuttling of the West Nile virus RNARNA ‐dependent RNARNA polymerase NS5NS5 is critical to infection

摘要