ZNF148 通过下调 RXRα 转录抑制 HBV 复制。

ZNF148 inhibits HBV replication by downregulating RXRα transcription.

机构信息

The Key Laboratory of Molecular Biology of Infectious Diseases designated by the Chinese Ministry of Education, Chongqing Medical University, Chong Yi Building, 1 YiXueYuan Road, Yuzhong District, Chongqing, 400016, China.

Department of Clinical Laboratory, Chongqing Traditional Chinese Medicine Hospital, Chongqing, China.

出版信息

Virol J. 2024 Jan 31;21(1):35. doi: 10.1186/s12985-024-02291-4.

DOI:10.1186/s12985-024-02291-4

PMID:38297280

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10832224/

Abstract

BACKGROUND

Progressive hepatitis B virus (HBV) infection can result in cirrhosis, hepatocellular cancer, and chronic hepatitis. While antiviral drugs that are now on the market are efficient in controlling HBV infection, finding a functional cure is still quite difficult. Identifying host factors involved in regulating the HBV life cycle will contribute to the development of new antiviral strategies. Zinc finger proteins have a significant function in HBV replication, according to earlier studies. Zinc finger protein 148 (ZNF148), a zinc finger transcription factor, regulates the expression of various genes by specifically binding to GC-rich sequences within promoter regions. The function of ZNF148 in HBV replication was investigated in this study.

METHODS

HepG2-Na/taurocholate cotransporting polypeptide (HepG2-NTCP) cells and Huh7 cells were used to evaluate the function of ZNF148 in vitro. Northern blotting and real-time PCR were used to quantify the amount of viral RNA. Southern blotting and real-time PCR were used to quantify the amount of viral DNA. Viral protein levels were elevated, according to the Western blot results. Dual-luciferase reporter assays were used to examine the transcriptional activity of viral promoters. ZNF148's impact on HBV in vivo was investigated using an established rcccDNA mouse model.

RESULTS

ZNF148 overexpression significantly decreased the levels of HBV RNAs and HBV core DNA in HBV-infected HepG2-NTCP cells and Huh7 cells expressing prcccDNA. Silencing ZNF148 exhibited the opposite effects in both cell lines. Furthermore, ZNF148 inhibited the activity of HBV ENII/Cp and the transcriptional activity of cccDNA. Mechanistic studies revealed that ZNF148 attenuated retinoid X receptor alpha (RXRα) expression by binding to the RXRα promoter sequence. RXRα binding site mutation or RXRα overexpression abolished the suppressive effect of ZNF148 on HBV replication. The inhibitory effect of ZNF148 was also observed in the rcccDNA mouse model.

CONCLUSIONS

ZNF148 inhibited HBV replication by downregulating RXRα transcription. Our findings reveal that ZNF148 may be a new target for anti-HBV strategies.

摘要

背景

乙型肝炎病毒（HBV）的进展性感染可导致肝硬化、肝细胞癌和慢性肝炎。虽然目前市场上的抗病毒药物在控制HBV 感染方面非常有效，但要找到功能性治愈仍然相当困难。鉴定参与调节 HBV 生命周期的宿主因素将有助于开发新的抗病毒策略。早期研究表明，锌指蛋白在 HBV 复制中具有重要作用。锌指蛋白 148（ZNF148）是一种锌指转录因子，通过特异性结合启动子区域内富含 GC 的序列来调节各种基因的表达。本研究旨在研究 ZNF148 在 HBV 复制中的作用。

方法

体外使用 HepG2-Na/牛磺胆酸钠共转运多肽（HepG2-NTCP）细胞和 Huh7 细胞评估 ZNF148 的功能。采用Northern 印迹和实时 PCR 定量病毒 RNA 量，采用 Southern 印迹和实时 PCR 定量病毒 DNA 量。根据 Western blot 结果，提高病毒蛋白水平。采用双荧光素酶报告基因检测试剂盒检测病毒启动子的转录活性。采用已建立的 rcccDNA 小鼠模型研究 ZNF148 对 HBV 的体内影响。

结果

ZNF148 过表达可显著降低 HBV 感染的 HepG2-NTCP 细胞和表达 prcccDNA 的 Huh7 细胞中 HBV RNAs 和 HBV 核心 DNA 的水平。在这两种细胞系中，沉默 ZNF148 则表现出相反的效果。此外，ZNF148 抑制 HBV ENII/Cp 的活性和 cccDNA 的转录活性。机制研究表明，ZNF148 通过结合 RXRα 启动子序列来抑制视黄醇 X 受体 α（RXRα）的表达。RXRα 结合位点突变或 RXRα 过表达可消除 ZNF148 对 HBV 复制的抑制作用。在 rcccDNA 小鼠模型中也观察到 ZNF148 的抑制作用。