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cccDNA Surrogate MC-HBV–Based Screen Identifies Cohesin Complex as a Novel HBV Restriction Factor
BACKGROUND & AIMS: Covalently closed circular DNA (cccDNA) of hepatitis B virus (HBV), existing as a stable minichromosome in the hepatocyte, is responsible for persistent HBV infection. Maintenance and sustained replication of cccDNA require its interaction with both viral and host proteins. Ho...
Autores principales: | , , , , , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9579331/ https://www.ncbi.nlm.nih.gov/pubmed/35987451 http://dx.doi.org/10.1016/j.jcmgh.2022.08.002 |
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author | Wu, Zhuanchang Wang, Liyuan Wang, Xin Sun, Yang Li, Haoran Zhang, Zhaoying Ren, Caiyue Zhang, Xiaohui Li, Shuangjie Lu, Jinghui Xu, Leiqi Yue, Xuetian Hong, Yue Li, Qiang Zhu, Haizhen Gong, Yaoqin Gao, Chengjiang Hu, Huili Gao, Lifen Liang, Xiaohong Ma, Chunhong |
author_facet | Wu, Zhuanchang Wang, Liyuan Wang, Xin Sun, Yang Li, Haoran Zhang, Zhaoying Ren, Caiyue Zhang, Xiaohui Li, Shuangjie Lu, Jinghui Xu, Leiqi Yue, Xuetian Hong, Yue Li, Qiang Zhu, Haizhen Gong, Yaoqin Gao, Chengjiang Hu, Huili Gao, Lifen Liang, Xiaohong Ma, Chunhong |
author_sort | Wu, Zhuanchang |
collection | PubMed |
description | BACKGROUND & AIMS: Covalently closed circular DNA (cccDNA) of hepatitis B virus (HBV), existing as a stable minichromosome in the hepatocyte, is responsible for persistent HBV infection. Maintenance and sustained replication of cccDNA require its interaction with both viral and host proteins. However, the cccDNA-interacting host factors that limit HBV replication remain elusive. METHODS: Minicircle HBV (MC-HBV), a recombinant cccDNA, was constructed based on chimeric intron and minicircle DNA technology. By mass spectrometry based on pull-down with biotinylated MC-HBV, the cccDNA-hepatocyte interaction profile was mapped. HBV replication was assessed in different cell models that support cccDNA formation. RESULTS: MC-HBV supports persistent HBV replication and mimics the cccDNA minichromosome. The MC-HBV–based screen identified cohesin complex as a cccDNA binding host factor, leading to reduced HBV replication. Mechanistically, with the help of CCCTC-binding factor (CTCF), which has specific binding sites on cccDNA, cohesin loads on cccDNA and reshapes cccDNA confirmation to prevent RNA polymerase II enrichment. Interestingly, HBV X protein transcriptionally reduces structural maintenance of chromosomes complex expression to partially relieve the inhibitory role of the cohesin complex on HBV replication. CONCLUSIONS: Our data not only provide a feasible approach to explore cccDNA-binding factors, but also identify cohesin/CTCF complex as a critical host restriction factor for cccDNA-driven HBV replication. These findings provide a novel insight into cccDNA–host interaction and targeted therapeutic intervention for HBV infection. |
format | Online Article Text |
id | pubmed-9579331 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-95793312022-10-20 cccDNA Surrogate MC-HBV–Based Screen Identifies Cohesin Complex as a Novel HBV Restriction Factor Wu, Zhuanchang Wang, Liyuan Wang, Xin Sun, Yang Li, Haoran Zhang, Zhaoying Ren, Caiyue Zhang, Xiaohui Li, Shuangjie Lu, Jinghui Xu, Leiqi Yue, Xuetian Hong, Yue Li, Qiang Zhu, Haizhen Gong, Yaoqin Gao, Chengjiang Hu, Huili Gao, Lifen Liang, Xiaohong Ma, Chunhong Cell Mol Gastroenterol Hepatol Original Research BACKGROUND & AIMS: Covalently closed circular DNA (cccDNA) of hepatitis B virus (HBV), existing as a stable minichromosome in the hepatocyte, is responsible for persistent HBV infection. Maintenance and sustained replication of cccDNA require its interaction with both viral and host proteins. However, the cccDNA-interacting host factors that limit HBV replication remain elusive. METHODS: Minicircle HBV (MC-HBV), a recombinant cccDNA, was constructed based on chimeric intron and minicircle DNA technology. By mass spectrometry based on pull-down with biotinylated MC-HBV, the cccDNA-hepatocyte interaction profile was mapped. HBV replication was assessed in different cell models that support cccDNA formation. RESULTS: MC-HBV supports persistent HBV replication and mimics the cccDNA minichromosome. The MC-HBV–based screen identified cohesin complex as a cccDNA binding host factor, leading to reduced HBV replication. Mechanistically, with the help of CCCTC-binding factor (CTCF), which has specific binding sites on cccDNA, cohesin loads on cccDNA and reshapes cccDNA confirmation to prevent RNA polymerase II enrichment. Interestingly, HBV X protein transcriptionally reduces structural maintenance of chromosomes complex expression to partially relieve the inhibitory role of the cohesin complex on HBV replication. CONCLUSIONS: Our data not only provide a feasible approach to explore cccDNA-binding factors, but also identify cohesin/CTCF complex as a critical host restriction factor for cccDNA-driven HBV replication. These findings provide a novel insight into cccDNA–host interaction and targeted therapeutic intervention for HBV infection. Elsevier 2022-08-17 /pmc/articles/PMC9579331/ /pubmed/35987451 http://dx.doi.org/10.1016/j.jcmgh.2022.08.002 Text en © 2022 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Original Research Wu, Zhuanchang Wang, Liyuan Wang, Xin Sun, Yang Li, Haoran Zhang, Zhaoying Ren, Caiyue Zhang, Xiaohui Li, Shuangjie Lu, Jinghui Xu, Leiqi Yue, Xuetian Hong, Yue Li, Qiang Zhu, Haizhen Gong, Yaoqin Gao, Chengjiang Hu, Huili Gao, Lifen Liang, Xiaohong Ma, Chunhong cccDNA Surrogate MC-HBV–Based Screen Identifies Cohesin Complex as a Novel HBV Restriction Factor |
title | cccDNA Surrogate MC-HBV–Based Screen Identifies Cohesin Complex as a Novel HBV Restriction Factor |
title_full | cccDNA Surrogate MC-HBV–Based Screen Identifies Cohesin Complex as a Novel HBV Restriction Factor |
title_fullStr | cccDNA Surrogate MC-HBV–Based Screen Identifies Cohesin Complex as a Novel HBV Restriction Factor |
title_full_unstemmed | cccDNA Surrogate MC-HBV–Based Screen Identifies Cohesin Complex as a Novel HBV Restriction Factor |
title_short | cccDNA Surrogate MC-HBV–Based Screen Identifies Cohesin Complex as a Novel HBV Restriction Factor |
title_sort | cccdna surrogate mc-hbv–based screen identifies cohesin complex as a novel hbv restriction factor |
topic | Original Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9579331/ https://www.ncbi.nlm.nih.gov/pubmed/35987451 http://dx.doi.org/10.1016/j.jcmgh.2022.08.002 |
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