Architecture and self-assembly of theSARS-CoV-2 nucleocapsid protein
Architecture and self-assembly of theSARS-CoV-2 nucleocapsid protein
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DOI：
10.1002/pro.3909
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发表时间：
2020-08-06
期刊：
影响因子：
8
通讯作者：
Corbett, Kevin D.
中科院分区：
文献类型：
--
作者：
Ye, Qiaozhen;West, Alan M. V.;Corbett, Kevin D.
2019年新型冠状病毒病大流行是二十一世纪世纪最严重的急性公共卫生威胁。为了通过强有力的测试和新的治疗方法来妥善解决这一危机，我们需要深入了解病原体SARS-CoV-2冠状病毒的生命周期。在这里，我们研究了SARS-CoV-2核衣壳蛋白的结构和自组装特性，该蛋白将病毒RNA包装成新的病毒体。我们确定了这种蛋白质的N2 b结构域的1.4埃分辨率晶体结构，揭示了一个紧凑的，相互缠绕的二聚体，类似于包括SARS-CoV在内的相关冠状病毒。当N2 B结构域在溶液中形成二聚体时，添加C-末端间隔区B/N3结构域介导同源四聚体的形成。使用氢-氘交换质谱法，我们发现的证据表明，至少有一部分的这种purone无序结构域的结构，可能形成一个α-螺旋，自关联和合作的N2 b域介导的四聚体形成。最后，我们从超过38，000个SARS-CoV-2基因组序列中绘制了N蛋白中氨基酸取代的位置。我们发现，这些取代强烈聚集在蛋白质的N2 a连接结构域中，并且N1 b和N2 b结构域内的取代远离其功能性RNA结合和二聚化界面聚集。总的来说，这项工作揭示了SARS-CoV-2生命周期中关键蛋白质的结构和自组装特性，对药物设计和基于抗体的测试都有影响。
The COVID-2019 pandemic is the most severe acute public health threat of the twenty-first century. To properly address this crisis with both robust testing and novel treatments, we require a deep understanding of the life cycle of the causative agent, the SARS-CoV-2 coronavirus. Here, we examine the architecture and self-assembly properties of the SARS-CoV-2 nucleocapsid protein, which packages viral RNA into new virions. We determined a 1.4 angstrom resolution crystal structure of this protein's N2b domain, revealing a compact, intertwined dimer similar to that of related coronaviruses including SARS-CoV. While the N2b domain forms a dimer in solution, addition of the C-terminal spacer B/N3 domain mediates formation of a homotetramer. Using hydrogen-deuterium exchange mass spectrometry, we find evidence that at least part of this putatively disordered domain is structured, potentially forming an alpha-helix that self-associates and cooperates with the N2b domain to mediate tetramer formation. Finally, we map the locations of amino acid substitutions in the N protein from over 38,000 SARS-CoV-2 genome sequences. We find that these substitutions are strongly clustered in the protein's N2a linker domain, and that substitutions within the N1b and N2b domains cluster away from their functional RNA binding and dimerization interfaces. Overall, this work reveals the architecture and self-assembly properties of a key protein in the SARS-CoV-2 life cycle, with implications for both drug design and antibody-based testing.