Coronavirus nucleocapsid proteins assemble constitutively in high molecular oligomers.

Coronavirus nucleocapsid proteins assemble constitutively in high molecular oligomers.
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DOI:
10.1038/s41598-017-06062-w
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发表时间:
2017-07-18
期刊:
影响因子:
4.6
通讯作者:
Reggiori F
Reggiori F
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Cong Y;Kriegenburg F;de Haan CAM;Reggiori F

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冠状病毒(CoV)是一种包膜病毒,依靠其核衣壳N蛋白将正链基因组RNA整合到病毒粒子中。冠状病毒N蛋白形成低聚体,但其多聚化的机制和相关性仍未完全了解。利用体外下拉实验和密度甘油梯度实验,我们发现至少有3个区域分布在其全长上,介导小鼠肝炎病毒(MHV)和严重急性呼吸综合征冠状病毒(SARS-CoV)N蛋白的自我相互作用。这些区域之间可以相互结合的事实为N蛋白齐聚提供了可能的分子基础。有趣的是,MHV感染细胞的细胞质N分子通过一种不需要与基因组RNA结合的过程结构性地组装成低聚物。基于我们的数据,我们提出了一个模型,在这个模型中,结构性N蛋白齐聚允许通过呈现多个病毒RNA结合基序来将基因组RNA最佳地负载到核糖核蛋白复合体中。
Coronaviruses (CoV) are enveloped viruses and rely on their nucleocapsid N protein to incorporate the positive-stranded genomic RNA into the virions. CoV N proteins form oligomers but the mechanism and relevance underlying their multimerization remain to be fully understood. Using in vitro pull-down experiments and density glycerol gradients, we found that at least 3 regions distributed over its entire length mediate the self-interaction of mouse hepatitis virus (MHV) and severe acute respiratory syndrome coronavirus (SARS-CoV) N protein. The fact that these regions can bind reciprocally between themselves provides a possible molecular basis for N protein oligomerization. Interestingly, cytoplasmic N molecules of MHV-infected cells constitutively assemble into oligomers through a process that does not require binding to genomic RNA. Based on our data, we propose a model where constitutive N protein oligomerization allows the optimal loading of the genomic viral RNA into a ribonucleoprotein complex via the presentation of multiple viral RNA binding motifs.
DOI: 10.1016/j.str.2005.08.021
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