RNA结合基序蛋白24（RBM24）通过介导HBV聚合酶和Epsilon元件之间

StephenW

J Virol. 2019 Jan 9. pii: JVI.02161-18. doi: 10.1128/JVI.02161-18. [Epub ahead of print]
RNA-Binding Motif Protein 24 (RBM24) Is Involved in pgRNA Packaging by Mediating Interaction between HBV Polymerase and the Epsilon Element.
Yao Y1,2, Yang B1,2, Chen Y3,4, Wang H3,4, Hu X3, Zhou Y3, Gao X5, Lu M6, Niu J5, Wen Z1, Wu C7, Chen X8.
Author information

1
    Joint Center of Translational Precision Medicine, Guangzhou Institute of Pediatrics, Guangzhou Women and Children Medical Center, Guangzhou, China.
2
    Joint Center of Translational Precision Medicine, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, China.
3
    Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, China.
4
    University of Chinese Academy of Sciences, Beijing, China.
5
    Department of Hepatology, The First Hospital of Jilin University, Changchun, Jilin, China.
6
    Institute of Virology, University Hospital of Essen, Essen, Germany.
7
    Joint Center of Translational Precision Medicine, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, China [email protected].
8
    Joint Center of Translational Precision Medicine, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, China [email protected].

Abstract

Encapsidation of pregenome RNA (pgRNA) is a curial step in HBV replication. Binding by viral polymerase (Pol) to the epsilon stem loop (ε) on the 5' terminal region (TR) of pgRNA is required for pgRNA packaging. However, the detailed mechanism is not well understood. RNA-binding motif protein 24 (RBM24) inhibits core translation by binding to the 5' TR of pgRNA. Here, we demonstrate that RBM24 is also involved in pgRNA packaging. RBM24 directly binds to the lower bulge of ε via RNA recognition submotifs (RNPs). RBM24 also interacts with Pol in an RNA-independent manner. The alanine-rich domain (ARD) of RBM24 and the reverse transcriptase domain (RT) of Pol are essential for binding between RBM24 and Pol. In addition, overexpression of RBM24 increases Pol-ε interaction, whereas RBM24 knockdown decreases interaction. RBM24 was able to rescue binding between ε and mutant Pol lacking ε-binding activity, further showing that RBM24 mediates interaction between Pol and ε by forming a Pol-RBM24-ε complex. Finally, RBM24 significantly promotes the packaging efficiency of pgRNA. In conclusion, RBM24 mediates Pol-ε interaction and formation of a Pol-RBM24-ε complex, which inhibits translation of pgRNA and results in pgRNA packing into capsids/virions for reverse transcription and DNA synthesis.IMPORTANCE Hepatitis B virus (HBV) is a ubiquitous human pathogen, and HBV infection is a major global health burden. Chronic HBV infection is associated with the development of liver diseases including fulminant hepatitis, hepatic fibrosis, cirrhosis, and hepatocellular carcinoma. A currently approved vaccine can prevent HBV infection, and medications are able to reduce viral load and prevent liver disease progression. However, current treatments rarely achieve a cure for chronic infection. Thus, it is important to gain insight into the mechanisms of HBV replication. In this study, we found that the host factor RBM24 is involved in pregenomic RNA (pgRNA) packaging and regulates HBV replication. These findings highlight a potential target for antiviral therapeutics of HBV infection.

Copyright © 2019 American Society for Microbiology.

PMID:
    30626666
DOI:
    10.1128/JVI.02161-18

StephenW

J Virol。 2019年1月9日.pii：JVI.02161-18。 doi：10.1128 / JVI.02161-18。 [提前打印]
RNA结合基序蛋白24（RBM24）通过介导HBV聚合酶和Epsilon元件之间的相互作用参与pgRNA包装。
Yao Y1,2，Yang B1,2，Chen Y3,4，Wang H3,4，Hu X3，Zhou Y3，Gao X5，Lu M6，Ni J5，Wen Z1，Wu C7，Chen X8。
作者信息

1
    广州市儿童医学中心广州儿科医学研究所转化精准医学联合中心，广州
2
    中国科学院武汉病毒研究所转化精准医学联合中心，武汉
3
    中国科学院武汉病毒研究所，武汉。
4
    中国科学院大学，北京，中国。
五
    吉林大学第一医院肝病科，吉林长春
6
    德国埃森埃森大学医院病毒学研究所。
7
    中国科学院武汉病毒研究所转化精准医学联合中心，武汉，中国武汉[email protected]。
8
    中国科学院武汉病毒研究所转化精准医学联合中心，中国武汉[email protected]。

抽象

前基因组RNA（pgRNA）的包装是HBV复制中的一个重要步骤。 pgRNA包装需要通过病毒聚合酶（Pol）与pgRNA的5'末端区域（TR）上的ε茎环（ε）结合。但是，详细的机制尚不清楚。 RNA结合基序蛋白24（RBM24）通过与pgRNA的5'TR结合来抑制核心翻译。在这里，我们证明RBM24也参与pgRNA包装。 RBM24通过RNA识别亚基因（RNP）直接与ε的下部凸起结合。 RBM24还以与RNA无关的方式与Pol相互作用。 RBM24的富含丙氨酸的结构域（ARD）和Pol的逆转录酶结构域（RT）对于RBM24和Pol之间的结合是必需的。此外，RBM24的过表达增加Pol-ε相互作用，而RBM24敲低降低相互作用。 RBM24能够拯救缺乏ε-结合活性的ε和突变体Pol之间的结合，进一步显示RBM24通过形成Pol-RBM24-ε复合物介导Pol和ε之间的相互作用。最后，RBM24显着提高了pgRNA的包装效率。总之，RBM24介导Pol-ε相互作用和Pol-RBM24-ε复合物的形成，其抑制pgRNA的翻译并导致pgRNA包装成衣壳/病毒体以进行逆转录和DNA合成。重要性乙型肝炎病毒（HBV）是一种无处不在的人类病原体和HBV感染是全球主要的健康负担。慢性HBV感染与肝脏疾病的发展有关，包括暴发性肝炎，肝纤维化，肝硬化和肝细胞癌。目前批准的疫苗可预防HBV感染，并且药物能够减少病毒载量并预防肝病进展。然而，目前的治疗很少能治愈慢性感染。因此，深入了解HBV复制的机制非常重要。在本研究中，我们发现宿主因子RBM24参与前基因组RNA（pgRNA）包装并调节HBV复制。这些发现突出了HBV感染的抗病毒治疗的潜在目标。

版权所有©2019美国微生物学会。

结论：
    30626666
DOI：
    10.1128 / JVI.02161-18