Grupo de Virología, Universidad El Bosque, Bogotá 110121, Colombia.
Viruses. 2023 Feb 9;15(2):481. doi: 10.3390/v15020481.
Ubiquitination and deubiquitination processes are widely involved in modulating the function, activity, localization, and stability of multiple cellular proteins regulating almost every aspect of cellular function. Several virus families have been shown to exploit the cellular ubiquitin-conjugating system to achieve a productive infection: enter the cell, promote genome replication, or assemble and release viral progeny. In this study, we analyzed the role of deubiquitinating enzymes (DUBs) during chikungunya virus (CHIKV) infection. HEK293T, Vero-E6, and Huh-7 cells were treated with two DUB inhibitors (PR619 or WP1130). Then, infected cells were evaluated by flow cytometry, and viral progeny was quantified using the plaque assay method. The changes in viral proteins and viral RNA were analyzed using Western blotting and RT-qPCR, respectively. Results indicate that treatment with DUB inhibitors impairs CHIKV replication due to significant protein and viral RNA synthesis deregulation. Therefore, DUB activity may be a pharmacological target for blocking CHIKV infection.
泛素化和去泛素化过程广泛参与调节多种细胞蛋白的功能、活性、定位和稳定性，这些蛋白调节着细胞功能的几乎各个方面。已经有几种病毒家族被证明利用细胞泛素连接系统来实现有效的感染：进入细胞、促进基因组复制或组装和释放病毒后代。在这项研究中，我们分析了去泛素化酶（DUB）在基孔肯雅病毒（CHIKV）感染过程中的作用。用两种 DUB 抑制剂（PR619 或 WP1130）处理 HEK293T、Vero-E6 和 Huh-7 细胞。然后，通过流式细胞术评估感染细胞，并用噬斑法定量病毒后代。分别用 Western blot 和 RT-qPCR 分析病毒蛋白和病毒 RNA 的变化。结果表明，由于蛋白质和病毒 RNA 合成的明显失调，DUB 抑制剂的处理会损害 CHIKV 的复制。因此，DUB 活性可能是阻断 CHIKV 感染的药理学靶点。
