Development of leafhopper cell culture to trace the early infection process of a nucleorhabdovirus, rice yellow stunt virus, in insect vector cells.
Development of leafhopper cell culture to trace the early infection process of a nucleorhabdovirus, rice yellow stunt virus, in insect vector cells.
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开发叶蝉细胞培养物以追踪核弹状病毒、稻黄矮化病毒在昆虫载体细胞中的早期感染过程
DOI：
10.1186/s12985-018-0987-6
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发表时间：
2018-04-20
期刊：
影响因子：
4.8
通讯作者：
Zhang XF
中科院分区：
文献类型：
--
作者：
Wang H;Wang J;Xie Y;Fu Z;Wei T;Zhang XF
水稻黄矮病毒（Rice yellow stunt virus，RYSV）是弹状病毒科（Rhabdoviridae）核弹状病毒属（Nucleorhabdovirus）的一种，在20世纪60、70年代曾严重威胁我国水稻生产。这种病毒的生物学的基本方面，如蛋白质定位和在昆虫载体细胞感染期间RYSV病毒质的形成，在很大程度上是未开发的。的具体作用（S）的结构蛋白核蛋白（N）和磷蛋白（P）在装配的病毒质在RYSV感染昆虫vector.MethodsIn本研究中，我们采用连续叶蝉细胞培养，免疫细胞化学技术，和透射电镜调查的亚细胞分布的N和P在RYSV感染。采用GST pull-down和酵母双杂交技术研究了N和P的体外相互作用。采用dsRNA干扰技术研究了N和P在RYSV病毒质组装中的功能作用。瞬时表达的N和P足以在Sf 9细胞中形成VpLS。此外，在离体条件下，N/P、N/N和P/P的交互作用也得到了证实。更有趣的是，积累的RYSV显着减少时，N基因或P基因的转录被敲低的dsRNA treatment.ConclusionsIn总结，我们的研究结果表明，N和P是主要的病毒因子，负责病毒质的形成在RYSV感染的昆虫细胞。在RYSV感染昆虫载体的早期，N和P在细胞核中相互作用形成病毒质样结构，这是RYSV感染所必需的。
BackgroundIn China, the rice pathogenRice yellow stunt virus(RYSV), a member of the genusNucleorhabdovirusin the familyRhabdoviridae, was a severe threat to rice production during the1960s and1970s. Fundamental aspects of the biology of this virus such as protein localization and formation of the RYSV viroplasm during infection of insect vector cells are largely unexplored. The specific role(s) of the structural proteins nucleoprotein (N) and phosphoprotein (P) in the assembly of the viroplasm during RYSV infection in insect vector is also unclear.MethodsIn present study, we used continuous leafhopper cell culture, immunocytochemical techniques, and transmission electron microscopy to investigate the subcellular distributions of N and P during RYSV infection. Both GST pull-down assay and yeast two-hybrid assay were used to assess the in vitro interaction of N and P. The dsRNA interference assay was performed to study the functional roles of N and P in the assembly of RYSV viroplasm.ResultsHere we demonstrated that N and P colocalized in the nucleus of RYSV-infectedNephotettix cincticepscell and formed viroplasm-like structures (VpLSs). The transiently expressed N and P are sufficient to form VpLSs in the Sf9 cells. In addition, the interactions of N/P, N/N and P/P were confirmed in vitro. More interestingly, the accumulation of RYSV was significantly reduced when the transcription of N gene or P gene was knocked down by dsRNA treatment.ConclusionsIn summary, our results suggest that N and P are the main viral factors responsible for the formation of viroplasm in RYSV-infected insect cells. Early during RYSV infection in the insect vector, N and P interacted with each other in the nucleus to form viroplasm-like structures, which are essential for the infection of RYSV.
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影响因子：
2.7
作者：
Amarasinghe GK;Bào Y;Basler CF;Bavari S;Beer M;Bejerman N;Blasdell KR;Bochnowski A;Briese T;Bukreyev A;Calisher CH;Chandran K;Collins PL;Dietzgen RG;Dolnik O;Dürrwald R;Dye JM;Easton AJ;Ebihara H;Fang Q;Formenty P;Fouchier RAM;Ghedin E;Harding RM;Hewson R;Higgins CM;Hong J;Horie M;James AP;Jiāng D;Kobinger GP;Kondo H;Kurath G;Lamb RA;Lee B;Leroy EM;Li M;Maisner A;Mühlberger E;Netesov SV;Nowotny N;Patterson JL;Payne SL;Paweska JT;Pearson MN;Randall RE;Revill PA;Rima BK;Rota P;Rubbenstroth D;Schwemmle M;Smither SJ;Song Q;Stone DM;Takada A;Terregino C;Tesh RB;Tomonaga K;Tordo N;Towner JS;Vasilakis N;Volchkov VE;Wahl-Jensen V;Walker PJ;Wang B;Wang D;Wang F;Wang LF;Werren JH;Whitfield AE;Yan Z;Ye G;Kuhn JH
通讯作者：
Kuhn JH
开发昆虫载体细胞培养和 RNA 干扰系统以研究斐济病毒复制蛋白的功能作用
影响因子：
5.4
作者：
Jia, Dongsheng;Chen, Hongyan;Wei, Taiyun
通讯作者：
Wei, Taiyun
影响因子：
2.1
作者：
CREAMER, R
通讯作者：
CREAMER, R
影响因子：
5
作者：
Ghosh, Debasish;Brooks, Robert E.;Goodin, Michael M.
通讯作者：
Goodin, Michael M.
影响因子：
5.4
作者：
Huang, YW;Geng, YF;Fang, RX
通讯作者：
Fang, RX