Host casein kinase 1-mediated phosphorylation modulates phase separation of a rhabdovirus phosphoprotein and virus infection.
Host casein kinase 1-mediated phosphorylation modulates phase separation of a rhabdovirus phosphoprotein and virus infection.
复制标题
宿主酪蛋白激酶 1 介导的磷酸化调节弹状病毒磷蛋白和病毒感染的相分离。
DOI：
10.7554/elife.74884
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发表时间：
2022-02-22
期刊：
影响因子：
7.7
通讯作者：
Wang XB
中科院分区：
文献类型：
--
作者：
Fang XD;Gao Q;Zang Y;Qiao JH;Gao DM;Xu WY;Wang Y;Li D;Wang XB
液-液相分离（LLPS）在细胞无膜细胞器的形成中起着重要作用。然而，在负义RNA（NSR）病毒感染期间，宿主因素如何调节病毒蛋白的LLPS在很大程度上是未知的。在这里，我们使用大麦黄纹花叶病毒（BYSMV）作为模型，以证明调节宿主酪蛋白激酶1（CK 1）的相分离和NSR病毒感染。我们首次发现BYSMV磷蛋白（P）形成具有液体性质的球形颗粒，并在体内招募病毒核苷酸（N）和聚合酶（L）蛋白。此外，P形成的颗粒被拴在ER/肌动蛋白网络中进行运输和融合。BYSMV P在体外形成液滴，并掺入N蛋白和基因组RNA的5′末端。有趣的是，BYSMV P的相分离被宿主CK 1依赖的内在无序P蛋白区域的磷酸化抑制。遗传分析表明，BYSMV P非磷酸化突变体呈现凝聚相，促进了病毒质的形成和病毒的复制。而磷酸化模拟突变体则以弥散相状态存在。总的来说，我们的研究结果表明，宿主CK 1调节病毒P蛋白和病毒感染的相分离。
Liquid-liquid phase separation (LLPS) plays important roles in forming cellular membraneless organelles. However, how host factors regulate LLPS of viral proteins during negative-sense RNA (NSR) virus infection is largely unknown. Here, we used barley yellow striate mosaic virus (BYSMV) as a model to demonstrate regulation of host casein kinase 1 (CK1) in phase separation and infection of NSR viruses. We first found that the BYSMV phosphoprotein (P) formed spherical granules with liquid properties and recruited viral nucleotide (N) and polymerase (L) proteins in vivo. Moreover, the P-formed granules were tethered to the ER/actin network for trafficking and fusion. BYSMV P alone formed droplets and incorporated the N protein and the 5′ trailer of genomic RNA in vitro. Interestingly, phase separation of BYSMV P was inhibited by host CK1-dependent phosphorylation of an intrinsically disordered P protein region. Genetic assays demonstrated that the unphosphorylated mutant of BYSMV P exhibited condensed phase, which promoted viroplasm formation and virus replication. Whereas, the phosphorylation-mimic mutant existed in diffuse phase state for virus transcription. Collectively, our results demonstrate that host CK1 modulates phase separation of the viral P protein and virus infection.