Saw Wuan Geok, Pan Ankita, Subramanian Manimekalai Malathy Sony, Grüber Gerhard
Nanyang Technological University, School of Biological Sciences, 60 Nanyang Drive, 637551, Singapore.
Nanyang Technological University, School of Biological Sciences, 60 Nanyang Drive, 637551, Singapore.
Antiviral Res. 2017 May;141:73-90. doi: 10.1016/j.antiviral.2017.02.005. Epub 2017 Feb 12.
Zika virus (ZIKV) has emerged as a pathogen of major health concern. The virus relies on its non-structural protein 5 (NS5) including a methyl-transferase (MTase) and a RNA-dependent RNA polymerase (RdRp) for capping and synthesis of the viral RNA and the nonstructural protein 3 (NS3) with its protease and helicase domain for polyprotein possessing, unwinding dsRNA proceeding replication, and NTPase/RTPase activities. In this study we present for the first time insights into the overall structure of the entire French Polynesia ZIKV NS3 in solution. The protein is elongated and flexible in solution. Solution studies of the individual protease- and helicase domains show the compactness of the two monomeric enzymes as well as the contribution of the 10-residues linker region to the flexibility of the entire NS3. We show also the solution X-ray scattering data of the French Polynesia ZIKV NS5, which is dimeric in solution and switches to oligomers in a concentration-dependent manner. The solution shapes of the MTase and RdRp domains are described. The dimer arrangement of ZIKV NS5 is discussed in terms of its importance for MTase-RdRp communication and concerted interaction with its flexible and monomeric counterpart NS3 during viral replication and capping. The comparison of ZIKV NS3 and -NS5 solution data with the related DENV nonstructural proteins shed light into the similarities and diversities of these classes of enzymes. Finally, the effect of ATPase inhibitors to the enzymatic active ZIKV NS3 and the individual helicase are provided.
寨卡病毒（ZIKV）已成为一个主要的健康问题病原体。该病毒依靠其非结构蛋白5（NS5）（包括一个甲基转移酶（MTase）和一个RNA依赖性RNA聚合酶（RdRp））来进行病毒RNA的加帽和合成，以及依靠具有蛋白酶和解旋酶结构域的非结构蛋白3（NS3）来进行多蛋白的加工、解开双链RNA以进行复制以及NTPase/RTPase活性。在本研究中，我们首次展示了法属波利尼西亚ZIKV NS3在溶液中的整体结构的见解。该蛋白在溶液中呈细长且灵活的状态。对单个蛋白酶和解旋酶结构域的溶液研究表明了这两种单体酶的紧密性以及10个残基连接区对整个NS3灵活性的贡献。我们还展示了法属波利尼西亚ZIKV NS5的溶液X射线散射数据，该蛋白在溶液中为二聚体，并以浓度依赖的方式转变为寡聚体。描述了MTase和RdRp结构域的溶液形状。讨论了ZIKV NS5的二聚体排列对于MTase-RdRp通讯以及在病毒复制和加帽过程中与其灵活的单体对应物NS3的协同相互作用的重要性。将ZIKV NS3和-NS5的溶液数据与相关的登革热病毒非结构蛋白进行比较，揭示了这些酶类的相似性和差异性。最后，提供了ATPase抑制剂对具有酶活性的ZIKV NS3和单个解旋酶的影响。
