Department of Biology, University of Dayton, Dayton, OH, United States of America.
Department of Medical Biochemistry and Microbiology, Uppsala University, Uppsala, Uppsala, Sweden.
PeerJ. 2023 Feb 20;11:e14776. doi: 10.7717/peerj.14776. eCollection 2023.
CCCH-type zinc figure proteins (ZFP) are small cellular proteins that are structurally maintained by zinc ions. Zinc ions coordinate the protein structure in a tetrahedral geometry by binding to cystine-cystine or cysteines-histidine amino acids. ZFP's unique structure enables it to interact with a wide variety of molecules including RNA; thus, ZFP modulates several cellular processes including the host immune response and virus replication. CCCH-type ZFPs have shown their antiviral efficacy against several DNA and RNA viruses. However, their role in the human coronavirus is little explored. We hypothesized that ZFP36L1 also suppresses the human coronavirus. To test our hypothesis, we used OC43 human coronavirus (HCoV) strain in our study. We overexpressed and knockdown ZFP36L1 in HCT-8 cells using lentivirus transduction. Wild type, ZFP36L1 overexpressed, and ZFP36L1 knockdown cells were each infected with HCoV-OC43, and the virus titer in each cell line was measured over 96 hours post-infection (p.i.). Our results show that HCoV-OC43 replication was significantly reduced with ZFP36L1 overexpression while ZFP36L1 knockdown significantly enhanced virus replication. ZFP36L1 knockdown HCT-8 cells started producing infectious virus at 48 hours p.i. which was an earlier timepoint as compared to wild -type and ZFP36L1 overexpressed cells. Wild-type and ZFP36L1 overexpressed HCT-8 cells started producing infectious virus at 72 hours p.i. Overall, the current study showed that overexpression of ZFP36L1 suppressed human coronavirus (OC43) production.
CCCH 型锌指蛋白（ZFP）是一种小型细胞蛋白，其结构由锌离子维持。锌离子通过与半胱氨酸-半胱氨酸或半胱氨酸-组氨酸氨基酸结合，以四面体几何形状配位蛋白质结构。ZFP 的独特结构使其能够与包括 RNA 在内的多种分子相互作用；因此，ZFP 调节包括宿主免疫反应和病毒复制在内的多种细胞过程。CCCH 型 ZFP 已显示出对几种 DNA 和 RNA 病毒的抗病毒功效。然而，它们在人类冠状病毒中的作用尚未得到充分探索。我们假设 ZFP36L1 也抑制人类冠状病毒。为了验证我们的假设，我们在研究中使用了 OC43 人类冠状病毒（HCoV）株。我们使用慢病毒转导在 HCT-8 细胞中过表达和敲低 ZFP36L1。野生型、ZFP36L1 过表达和 ZFP36L1 敲低细胞分别感染 HCoV-OC43，感染后 96 小时（p.i.）测量每个细胞系中的病毒滴度。我们的结果表明，ZFP36L1 过表达显著降低了 HCoV-OC43 的复制，而 ZFP36L1 敲低则显著增强了病毒复制。ZFP36L1 敲低的 HCT-8 细胞在感染后 48 小时开始产生感染性病毒，这比野生型和 ZFP36L1 过表达细胞更早。野生型和 ZFP36L1 过表达的 HCT-8 细胞在感染后 72 小时开始产生感染性病毒。总体而言，本研究表明 ZFP36L1 的过表达抑制了人类冠状病毒（OC43）的产生。
