Double-stranded RNA drives SARS-CoV-2 nucleocapsid protein to undergo phase separation at specific temperatures.
Double-stranded RNA drives SARS-CoV-2 nucleocapsid protein to undergo phase separation at specific temperatures.
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DOI：
10.1093/nar/gkac596
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发表时间：
2022-08-12
影响因子：
14.9
通讯作者：
Gladfelter, Amy S.
中科院分区：
文献类型：
--
作者：
Roden, Christine A.;Dai, Yifan;Giannetti, Catherine A.;Seim, Ian;Lee, Myungwoon;Sealfon, Rachel;McLaughlin, Grace A.;Boerneke, Mark A.;Iserman, Christiane;Wey, Samuel A.;Ekena, Joanne L.;Troyanskaya, Olga G.;Weeks, Kevin M.;You, Lingchong;Chilkoti, Ashutosh;Gladfelter, Amy S.
核衣壳蛋白（n蛋白）在冠状病毒复制的多个步骤中都是必需的。sars - cov -2- n蛋白在特定温度下与特定病毒RNA冷凝，使其成为破译冷凝物中RNA序列特异性的强大模型。我们鉴定了两个独立的和不同的双链RNA基序（dsRNA贴纸），促进n蛋白凝聚。这些dsRNA贴纸分别被n蛋白的两个RNA结合域（rbd）识别。RBD1倾向于具有转录调控序列（TRS）等序列的结构化RNA。RBD2倾向于独立于序列的长片段dsRNA。因此，两种n蛋白rbd与不同的dsRNA贴纸相互作用，这些相互作用赋予了特定的液滴物理特性，可以支持各种病毒功能。具体来说，我们发现dsRNA的加入降低了依赖于RBD2相互作用的冷凝温度，并调节了翻译抑制。相反，RBD1位点是亚基因组（sg） RNA生成和促进gRNA压缩的关键序列。TRS-L/B序列附近RBD1结合基序的密度与亚基因组RNA生成水平相关。向包装的转变可能是由RBD1相互作用介导的，RBD1相互作用产生的颗粒再现了病毒粒子的包装单元。因此，SARS-CoV-2可以实现生化复杂性，在相同的细胞质中执行多种功能，以最小的蛋白质成分为基础，利用控制n蛋白相互作用的多个不同RNA基序。
Nucleocapsid protein (N-protein) is required for multiple steps in betacoronaviruses replication. SARS-CoV-2-N-protein condenses with specific viral RNAs at particular temperatures making it a powerful model for deciphering RNA sequence specificity in condensates. We identify two separate and distinct double-stranded, RNA motifs (dsRNA stickers) that promote N-protein condensation. These dsRNA stickers are separately recognized by N-protein's two RNA binding domains (RBDs). RBD1 prefers structured RNA with sequences like the transcription-regulatory sequence (TRS). RBD2 prefers long stretches of dsRNA, independent of sequence. Thus, the two N-protein RBDs interact with distinct dsRNA stickers, and these interactions impart specific droplet physical properties that could support varied viral functions. Specifically, we find that addition of dsRNA lowers the condensation temperature dependent on RBD2 interactions and tunes translational repression. In contrast RBD1 sites are sequences critical for sub-genomic (sg) RNA generation and promote gRNA compression. The density of RBD1 binding motifs in proximity to TRS-L/B sequences is associated with levels of sub-genomic RNA generation. The switch to packaging is likely mediated by RBD1 interactions which generate particles that recapitulate the packaging unit of the virion. Thus, SARS-CoV-2 can achieve biochemical complexity, performing multiple functions in the same cytoplasm, with minimal protein components based on utilizing multiple distinct RNA motifs that control N-protein interactions.
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影响因子：
9.8
作者：
Jack A;Ferro LS;Trnka MJ;Wehri E;Nadgir A;Nguyenla X;Fox D;Costa K;Stanley S;Schaletzky J;Yildiz A
通讯作者：
Yildiz A
病毒粒子内 SARS-CoV-2 RNA 基因组的结构
影响因子：
16.6
作者：
Cao C;Cai Z;Xiao X;Rao J;Chen J;Hu N;Yang M;Xing X;Wang Y;Li M;Zhou B;Wang X;Wang J;Xue Y
通讯作者：
Xue Y
影响因子：
16
作者：
Huston NC;Wan H;Strine MS;de Cesaris Araujo Tavares R;Wilen CB;Pyle AM
通讯作者：
Pyle AM
影响因子：
4.3
作者：
Choi, Jeong-Mo;Dar, Furqan;Pappu, Rohit, V
通讯作者：
Pappu, Rohit, V
影响因子：
7.6
作者：
Chang CK;Hou MH;Chang CF;Hsiao CD;Huang TH
通讯作者：
Huang TH