SARS-CoV-2 nucleocapsid protein phase-separates with RNA and with human hnRNPs.
SARS-CoV-2 nucleocapsid protein phase-separates with RNA and with human hnRNPs.
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DOI：
10.15252/embj.2020106478
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发表时间：
2020-12-15
期刊：
影响因子：
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通讯作者：
Fawzi NL
中科院分区：
文献类型：
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作者：
Perdikari TM;Murthy AC;Ryan VH;Watters S;Naik MT;Fawzi NL
核衣壳蛋白和基因组RNA的紧密包装复合物形成病毒的核心，并在病毒工厂内组装，病毒工厂是在与人类应激颗粒相关的宿主细胞内形成的动态隔室。在这里，我们测试了来自严重急性呼吸综合征冠状病毒2（SARS-CoV-2）的多价RNA结合核衣壳蛋白（N）通过液-液相分离（LLPS）与RNA缩合的可能性，以及N蛋白可以在与SG形成相关的人类RNA结合蛋白的相分离形式中募集。具有RNA的稳健LLPS需要两个固有无序区（IDR），N末端IDR和中心接头IDR，以及折叠的C末端寡聚化结构域，而折叠的N末端结构域和C末端IDR不是必需的。通过添加非特异性RNA诱导N蛋白相分离。此外，N在体外分成相分离形式的全长人hnRNP（TDP-43、FUS、hnRNPA 2）及其低复杂性结构域（LC）。这些结果为N在SARS-CoV-2病毒基因组包装和病毒复制和感染所需的宿主蛋白质选择中的作用提供了潜在的机制。来自SARS-CoV-2的核衣壳蛋白与RNA进行液-液相分离，并可分配到由TDP-43、FUS或hnRNPA 2形成的液滴中。
Tightly packed complexes of nucleocapsid protein and genomic RNA form the core of viruses and assemble within viral factories, dynamic compartments formed within the host cells associated with human stress granules. Here, we test the possibility that the multivalent RNA‐binding nucleocapsid protein (N) from severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) condenses with RNA via liquid–liquid phase separation (LLPS) and that N protein can be recruited in phase‐separated forms of human RNA‐binding proteins associated with SG formation. Robust LLPS with RNA requires two intrinsically disordered regions (IDRs), the N‐terminal IDR and central‐linker IDR, as well as the folded C‐terminal oligomerization domain, while the folded N‐terminal domain and the C‐terminal IDR are not required. N protein phase separation is induced by addition of non‐specific RNA. In addition, N partitions in vitro into phase‐separated forms of full‐length human hnRNPs (TDP‐43, FUS, hnRNPA2) and their low‐complexity domains (LCs). These results provide a potential mechanism for the role of N in SARS‐CoV‐2 viral genome packing and in host‐protein co‐opting necessary for viral replication and infectivity. The nucleocapsid protein from SARS‐CoV‐2 undergoes liquid‐liquid phase separation with RNA and can partition into droplets formed by TDP‐43, FUS or hnRNPA2.