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Original Article | Open Access

CUL4B facilitates HBV replication by promoting HBx stabilization

Haixia Shan1,2,*, Bo Wang1,3,*, Xiaodong Zhang1,*, Hui Song1, Xi Li4, Yongxin Zou4, Baichun Jiang4, Huili Hu4, Hao Dou4, Changshun Shao4, Lifen Gao1,5, Chunhong Ma1,5, Xiaoyun Yang6 ( ), Xiaohong Liang1,5 ( ), Yaoqin Gong4 ( )
Key Laboratory for Experimental Teratology of the Ministry of Education, Key Laboratory of Infection and Immunity of Shandong Province and Department of Immunology, School of Basic Medical Sciences, Shandong University, Jinan 250012, China
Department of Laboratory Diagnosis, Cangzhou Combination of Traditional Chinese and Western Medicine Hospital of Hebei Province, Cangzhou 061000, China
Division of Gastroenterology and Hepatology, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai Institute of Digestive Disease, Shanghai 200120, China
Ministry of Education Key Laboratory of Experimental Teratology and Institute of Molecular Medicine and Genetics, School of Basic Medical Sciences, Shandong University, Jinan 250012, China
Collaborative Innovation Center of Technology and Equipment for Biological Diagnosis and Therapy in Universities of Shandong, Jinan 250012, China
Department of Gastroenterology, Qilu Hospital, Shandong University, Jinan 250012, China

*These authors contributed equally to this work.

Show Author Information

Abstract

Objective

Hepatitis B virus (HBV) infection is a major public health problem worldwide. However, the regulatory mechanisms underlying HBV replication remain unclear. Cullin 4B-RING ubiquitin E3 ligase (CRL4B) is involved in regulating diverse physiological and pathophysiological processes. In our study, we aimed to explain the role of CUL4B in HBV infection.

Methods

Cul4b transgenic mice or conditional knockout mice, as well as liver cell lines with CUL4B overexpression or knockdown, were used to assess the role of CUL4B in HBV replication. Immunoprecipitation assays and immunofluorescence staining were performed to study the interaction between CUL4B and HBx. Cycloheximide chase assays and in vivo ubiquitination assays were performed to evaluate the half-life and the ubiquitination status of HBx.

Results

The hydrodynamics-based hepatitis B model in Cul4b transgenic or conditional knockout mice indicated that CUL4B promoted HBV replication (P < 0.05). Moreover, the overexpression or knockdown system in human liver cell lines validated that CUL4B increased HBV replication in an HBx-dependent manner. Importantly, immunoprecipitation assays and immunofluorescence staining showed an interaction between CUL4B and HBx. Furthermore, CUL4B upregulated HBx protein levels by inhibiting HBx ubiquitination and proteasomal degradation (P < 0.05). Finally, a positive correlation between CUL4B expression and HBV pgRNA level was observed in liver tissues from HBV-positive patients and HBV transgenic mice.

Conclusions

CUL4B enhances HBV replication by interacting with HBx and disrupting its ubiquitin-dependent proteasomal degradation. CUL4B may therefore be a potential target for anti-HBV therapy.

Keywords

HBVHCCubiquitinationCUL4BHBx

Electronic Supplementary Material

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Cancer Biology & Medicine
Volume 19 Issue 1,
January 2022
Pages 120-131
DOI: 10.20892/j.issn.2095-3941.2020.0468
Cite this article:
Shan H, Wang B, Zhang X, et al. CUL4B facilitates HBV replication by promoting HBx stabilization. Cancer Biology & Medicine, 2022, 19(1): 120-131. https://doi.org/10.20892/j.issn.2095-3941.2020.0468

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Received: 12 August 2020
Accepted: 24 December 2020
Published: 21 March 2022
©2022 Cancer Biology & Medicine.

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