Maxwell H. Gluck Equine Research Center, Department of Veterinary Science, University of Kentucky, Lexington, Kentucky, USA.
Department of Biology and Microbiology, South Dakota State University, Brookings, South Dakota, USA.
J Med Virol. 2022 Nov;94(11):5392-5400. doi: 10.1002/jmv.27991. Epub 2022 Jul 23.
The global spread of the mosquito-borne Zika virus (ZIKV) infection and its complications including Guillain-Barré syndrome and fetus microcephaly in 2015 have made ZIKV as a significant public health threat. The capsid protein plays crucial roles in ZIKV replication and thus represents an attractive therapeutic target. However, inhibitors of ZIKV capsid assembly have not been rigorously identified due to the lack of a target-based screening system. In this study, we developed a novel ZIKV capsid interaction method based on a split-luciferase complementation assay, which can be used to measure and quantify ZIKV capsid-capsid (C-C) interaction by the restored luciferase signal when capsid proteins interact with each other. Furthermore, a Tet-on inducible stable cell line was generated to screen inhibitors of capsid dimerization. By using of this system, peptides (Pep.15-24 in the N-terminal region of ZIKV capsid protein and Pep.44-58 in the α2 helix of ZIKV capsid protein) were identified to inhibit ZIKV C-C interaction. Overall, this study developed a novel inducible assay system to measure ZIKV capsid interaction and identify ZIKV capsid multimerization inhibitors, which will be applied for future discovery of ZIKV assembly inhibitors.
2015 年，蚊媒传播的 Zika 病毒（ZIKV）感染在全球范围内的传播及其并发症，包括格林-巴利综合征和胎儿小头畸形，使 ZIKV 成为一个重大的公共卫生威胁。衣壳蛋白在 ZIKV 复制中起着至关重要的作用，因此它是一个有吸引力的治疗靶点。然而，由于缺乏基于靶点的筛选系统，ZIKV 衣壳组装抑制剂尚未得到严格鉴定。在这项研究中，我们开发了一种基于荧光素酶互补测定的新型 ZIKV 衣壳相互作用方法，该方法可通过衣壳蛋白相互作用时恢复的荧光素酶信号来测量和定量 ZIKV 衣壳-衣壳（C-C）相互作用。此外，还生成了 Tet-on 诱导的稳定细胞系，用于筛选衣壳二聚化抑制剂。通过使用该系统，鉴定出肽（ZIKV 衣壳蛋白 N 端区域的 Pep.15-24 和 ZIKV 衣壳蛋白 α2 螺旋的 Pep.44-58）可抑制 ZIKV C-C 相互作用。总的来说，这项研究开发了一种新的诱导测定系统来测量 ZIKV 衣壳相互作用，并鉴定 ZIKV 衣壳多聚化抑制剂，这将应用于未来 ZIKV 组装抑制剂的发现。
