Birnaviridae virus factories show features of liquid-liquid phase separation, and are distinct from paracrystalline arrays of virions observed by electron microscopy
Birnaviridae virus factories show features of liquid-liquid phase separation, and are distinct from paracrystalline arrays of virions observed by electron microscopy
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双RNA病毒科病毒工厂表现出液-液相分离的特征，与电子显微镜观察到的病毒颗粒的旁晶阵列不同
DOI：
10.1101/2021.11.16.468875
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发表时间：
2021
期刊：
影响因子：
--
通讯作者：
Reddy V
中科院分区：
文献类型：
--
作者：
Reddy V
为了获得更多关于双核糖核酸病毒工厂（VF）性质的信息，我们使用了一种重组传染性法氏囊病病毒（IBDV），该病毒表达了与聚合酶（VP 1）相连的分裂型GFP 11，我们之前已经证明该聚合酶是表达GFP 1 -10的感染细胞中VF的标记。我们发现，VF共定位与5-乙炔基尿苷在放线菌素的存在下，表明它们含有新合成的病毒RNA，和VF是可见的感染细胞，固定和透性与毛地黄皂苷，表明他们不是膜结合。荧光恢复后的光漂白（FRAP）的一个区域内的VF的兴趣迅速发生，恢复约25%至87%的原始强度超过146秒，和VF被溶解的1，6-己二醇处理，证明他们表现出的性能与液-液相分离一致。与VP 3（MC，0.9）相比，VF GFP信号与衣壳蛋白VP 2（Manders'系数[MC] 0.6）的共定位较低，这促使我们通过透射电子显微镜（TEM）研究VF超微结构。在感染的细胞中，在细胞质中观察到病毒粒子的准晶体阵列（PA），以及离散的电子致密区域。利用相关光学和电子显微镜（CLEM），我们观察到的电子致密区相关的GFP信号的VF，这是不同于PA。总之，双核糖核酸病毒VF含有新合成的病毒RNA，不被膜结合，显示出与液-液相分离一致的性质，并且与通过TEM观察到的PA不同。重要的是，双核糖核酸病毒的成员感染鸟类、鱼类和昆虫，并且是对家禽业和水产养殖具有重要经济意义的疾病的原因。尽管它们很重要，但它们如何在细胞中复制仍然知之甚少。在这里，我们表明，theNiraviridae病毒工厂是不膜结合的，表现出与液-液相分离相一致的特性，并且与透射电子显微镜观察到的病毒粒子的准晶体阵列不同，增强了我们对病毒复制的基础知识，这些知识可用于开发控制疾病的策略，或优化其治疗应用。
To gain more information about the nature ofBirnaviridaevirus factories (VFs), we used a recombinant infectious bursal disease virus (IBDV) expressing split-GFP11 tagged to the polymerase (VP1) that we have previously shown is a marker for VFs in infected cells expressing GFP1-10. We found that VFs colocalized with 5-ethynyl uridine in the presence of actinomycin, demonstrating they contained newly synthesized viral RNA, and VFs were visible in infected cells that were fixed and permeabilized with digitonin, demonstrating that they were not membrane bound. Fluorescence recovery after photobleaching (FRAP) a region of interest within the VFs occurred rapidly, recovering from approximately 25% to 87% the original intensity over 146 s, and VFs were dissolved by 1,6-hexanediol treatment, demonstrating they showed properties consistent with liquid-liquid phase separation. There was a lower colocalization of the VF GFP signal with the capsid protein VP2 (Manders’ coefficient [MC] 0.6), compared to VP3 (MC, 0.9), which prompted us to investigate the VF ultrastructure by transmission electron microscopy (TEM). In infected cells, paracrystalline arrays (PAs) of virions were observed in the cytoplasm, as well as discrete electron dense regions. Using correlative light and electron microscopy (CLEM), we observed that the electron dense regions correlated with the GFP signal of the VFs, which were distinct from the PAs. In summary,BirnaviridaeVFs contain newly synthesized viral RNA, are not bound by a membrane, show properties consistent with liquid-liquid phase separation, and are distinct from the PAs observed by TEM.IMPORTANCEMembers of theBirnaviridaeinfect birds, fish and insects, and are responsible for diseases of significant economic importance to the poultry industry and aquaculture. Despite their importance, how they replicate in cells remains poorly understood. Here, we show that theBirnaviridaevirus factories are not membrane bound, demonstrate properties consistent with liquid-liquid phase separation, and are distinct from the paracrystalline arrays of virions observed by transmission electron microscopy, enhancing our fundamental knowledge of virus replication that could be used to develop strategies to control disease, or optimize their therapeutic application.
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影响因子：
3.8
作者：
Delmas, Bernard;Attoui, Houssam;Vakharia, Vikram N.
通讯作者：
Vakharia, Vikram N.
传染性法氏囊病病毒蛋白的表达和加工。
DOI：
--
发表时间：
2000
期刊：
影响因子：
--
作者：
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Wark Kl
影响因子：
5.4
作者：
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Delmas, B
影响因子：
14.9
作者：
Koulouras G;Panagopoulos A;Rapsomaniki MA;Giakoumakis NN;Taraviras S;Lygerou Z
通讯作者：
Lygerou Z
DOI：
10.15252/embj.2021107711
发表时间：
2021-11-02
期刊：
The EMBO journal
影响因子：
--
作者：
Geiger F;Acker J;Papa G;Wang X;Arter WE;Saar KL;Erkamp NA;Qi R;Bravo JP;Strauss S;Krainer G;Burrone OR;Jungmann R;Knowles TP;Engelke H;Borodavka A
通讯作者：
Borodavka A