cccDNA Surrogate MC-HBV-Based Screen Identifies Cohesin Complex as a Novel HBV Restriction Factor.
cccDNA Surrogate MC-HBV-Based Screen Identifies Cohesin Complex as a Novel HBV Restriction Factor.
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基于 cccDNA 替代 MC-HBV 的筛选将 Cohesin 复合物鉴定为新型 HBV 限制因子。
DOI：
10.1016/j.jcmgh.2022.08.002
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发表时间：
2022
影响因子：
7.2
通讯作者：
Ma, Chunhong
中科院分区：
文献类型：
--
作者：
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
B型肝炎病毒（HBV）的互补闭合环状DNA（cccDNA）作为稳定的微小染色体存在于肝细胞中，是HBV持续感染的主要原因。cccDNA的维持和持续复制需要其与病毒和宿主蛋白的相互作用。然而，限制HBV复制的cccDNA相互作用的宿主因子仍然难以捉摸。基于嵌合内含子和小环DNA技术构建了重组cccDNA--小环HBV（MC-HBV）。通过基于用生物素化MC-HBV下拉的质谱法，绘制cccDNA-肝细胞相互作用谱。在支持cccDNA形成的不同细胞模型中评估HBV复制。MC-HBV支持持续的HBV复制并模拟cccDNA微型染色体。基于MC-HBV的筛选将粘附素复合物鉴定为cccDNA结合宿主因子，导致HBV复制减少。从机制上讲，在cccDNA上具有特异性结合位点的CCCTC结合因子（CTCF）的帮助下，粘附素加载在cccDNA上并重塑cccDNA构象，以防止RNA聚合酶II富集。有趣的是，HBV X蛋白转录减少染色体复合体表达的结构维持，以部分减轻粘附素复合体对HBV复制的抑制作用。我们的数据不仅提供了一个可行的方法来探索cccDNA结合因子，但也确定cohesin/CTCF复合物作为一个关键的宿主限制因子cccDNA驱动的HBV复制。这些发现为HBV感染的cccDNA-宿主相互作用和靶向治疗干预提供了新的见解。
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. 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. 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. 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.
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影响因子：
3.3
作者：
Gallego-Paez LM;Tanaka H;Bando M;Takahashi M;Nozaki N;Nakato R;Shirahige K;Hirota T
通讯作者：
Hirota T
影响因子：
30.3
作者：
Arvey A;Tempera I;Tsai K;Chen HS;Tikhmyanova N;Klichinsky M;Leslie C;Lieberman PM
通讯作者：
Lieberman PM
影响因子：
3
作者：
Bader, GD;Hogue, CW
通讯作者：
Hogue, CW
DOI：
10.1007/978-1-4939-6700-1_2
发表时间：
2017-01-01
期刊：
HEPATITIS B VIRUS: METHODS AND PROTOCOLS
影响因子：
--
作者：
Ni, Yi;Urban, Stephan
通讯作者：
Urban, Stephan
DOI：
10.1016/j.cub.2014.08.011
发表时间：
2014-10-06
期刊：
Current biology : CB
影响因子：
--
作者：
Ladurner R;Bhaskara V;Huis in 't Veld PJ;Davidson IF;Kreidl E;Petzold G;Peters JM
通讯作者：
Peters JM