Structural Dynamics of the C-terminal X Domain of Nipah and Hendra Viruses Controls the Attachment to the C-terminal Tail of the Nucleocapsid Protein
Structural Dynamics of the C-terminal X Domain of Nipah and Hendra Viruses Controls the Attachment to the C-terminal Tail of the Nucleocapsid Protein
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DOI：
10.1016/j.jmb.2022.167551
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发表时间：
2022-04-07
影响因子：
5.6
通讯作者：
Jamin, Marc
中科院分区：
文献类型：
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作者：
Bourhis, Jean-Marie;Yabukarski, Filip;Jamin, Marc
为了了解副粘病毒科病毒的转录/复制复合体中磷蛋白（P）和核蛋白（N）之间的动态相互作用，并解释它们在调节病毒增殖中的作用，我们表征了尼帕病毒（NiV）和亨德拉病毒（HeV）P蛋白C-末端X结构域（P-XD）的结构特性。在晶体中，分离的NiV P-XD采用双螺旋二聚体构象，这是不能够结合其合作伙伴，但在复杂的C-末端的N蛋白（N-TAIL）的内在无序的尾巴，它折叠成一个典型的3 H束构象。在溶液中，SEC-MALLS，SAXS和NMR光谱实验表明，NiV和HeV P-XD的尺寸大于相同分子量的紧凑蛋白质的预期尺寸，并且处于紧凑的三螺旋（3 H）束和部分未折叠构象之间的构象交换中，其中螺旋α（3）与其他两个分离。一些测量也提供了强有力的证据，在溶液中的NiV P-XD的二聚化，但不为HeV P-XD。实验SAXS数据和螺旋动力学模型的Enhancement建模调和所有这些数据，产生一个模型，其中NiV和HeV P-XD在不同构象之间交换，并且NiV而不是HeV P-XD形成二聚体。最后，拯救了包含携带HeV P-XD的嵌合P的重组NiV，并与亲本NiV进行比较。在细胞中进行的实验表明，P-XD的替代没有显着影响复制动力学，但引起轻微的病毒减毒，这表明NiV P-XD的二聚化在病毒复制中可能起作用。(C)2022爱思唯尔有限公司保留所有权利。
To understand the dynamic interactions between the phosphoprotein (P) and the nucleoprotein (N) within the transcription/replication complex of the Paramyxoviridae and to decipher their roles in regulating viral multiplication, we characterized the structural properties of the C-terminal X domain (P-XD) of Nipah (NiV) and Hendra virus (HeV) P protein. In crystals, isolated NiV P-XD adopted a two-helix dimeric conformation, which was incompetent for binding its partners, but in complex with the C-terminal intrinsically disordered tail of the N protein (N-TAIL), it folded into a canonical 3H bundle conformation. In solution, SEC-MALLS, SAXS and NMR spectroscopy experiments indicated that both NiV and HeV P-XD were larger in size than expected for compact proteins of the same molecular mass and were in conformational exchange between a compact three-helix (3H) bundle and partially unfolded conformations, where helix alpha(3) is detached from the other two. Some measurements also provided strong evidence for dimerization of NiV P-XD in solution but not for HeV P-XD. Ensemble modeling of experimental SAXS data and statistical-dynamical modeling reconciled all these data, yielding a model where NiV and HeV P-XD exchanged between different conformations, and where NiV but not HeV P-XD formed dimers. Finally, recombinant NiV comprising a chimeric P carrying HeV P-XD was rescued and compared with parental NiV. Experiments carried out in cellula demonstrated that the replacement of P-XD did not significantly affect the replication dynamics while caused a slight virus attenuation, suggesting a possible role of the dimerization of NiV P-XD in viral replication. (C) 2022 Elsevier Ltd. All rights reserved.