Measles virus nucleo- and phosphoproteins form liquid-like phase-separated compartments that promote nucleocapsid assembly
Measles virus nucleo- and phosphoproteins form liquid-like phase-separated compartments that promote nucleocapsid assembly
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DOI：
10.1126/sciadv.aaz7095
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发表时间：
2020-04-01
期刊：
影响因子：
13.6
通讯作者：
Blackledge, Martin
中科院分区：
文献类型：
--
作者：
Guseva, Serafima;Milles, Sigrid;Blackledge, Martin
已知许多病毒形成细胞隔间，也称为病毒工厂。副粘病毒，包括麻疹病毒，将其蛋白质组学和基因组物质共定位在受感染细胞的点状体中。我们证明纯化的麻疹病毒核蛋白(N)和磷酸化蛋白(P)在体外混合后形成液体样无膜细胞器。我们确定了涉及该过程中涉及的N和P的内在无序域的弱相互作用，其中一个是相分离所必需的。荧光使我们能够跟踪液滴形成时N和P的动力学调制，而核磁共振用于研究这一过程的热力学。RNA与液滴共定位，在液滴中触发N原体组装成包裹RNA的核衣壳样颗粒。与稀相相比，液滴内的包封率提高，揭示了液-液分离在麻疹病毒复制中的作用之一。
Many viruses are known to form cellular compartments, also called viral factories. Paramyxoviruses, including measles virus, colocalize their proteomic and genomic material in puncta in infected cells. We demonstrate that purified nucleoproteins (N) and phosphoproteins (P) of measles virus form liquid-like membraneless organelles upon mixing in vitro. We identify weak interactions involving intrinsically disordered domains of N and P that are implicated in this process, one of which is essential for phase separation. Fluorescence allows us to follow the modulation of the dynamics of N and P upon droplet formation, while NMR is used to investigate the thermodynamics of this process. RNA colocalizes to droplets, where it triggers assembly of N protomers into nucleocapsid-like particles that encapsidate the RNA. The rate of encapsidation within droplets is enhanced compared to the dilute phase, revealing one of the roles of liquid-liquid phase separation in measles virus replication.