跨抗原譜分析揭示了在HBV感染過程中新型的自我限制性病毒-

StephenW

Translatomic profiling reveals novel self-restricting virus-host interactions during HBV infection
Shilin Yuan  1 , Guanghong Liao  2 , Menghuan Zhang  3 , Yuanfei Zhu  4 , Kun Wang  5 , Weidi Xiao  6 , Caiwei Jia  7 , Minhui Dong  8 , Na Sun  9 , Axel Walch  10 , Ping Xu  11 , Jiming Zhang  12 , Qiang Deng  13 , Ronggui Hu  14
Affiliations
Affiliations

    1
    State Key Laboratory of Molecular Biology, Shanghai Institute of Biochemistry and Cell Biology, Center for Excellence in Molecular Cell Science, Chinese Academy of Sciences, Shanghai 200031, China; University of Chinese Academy of Sciences, Beijing 100049, China. Electronic address: [email protected].
    2
    State Key Laboratory of Molecular Biology, Shanghai Institute of Biochemistry and Cell Biology, Center for Excellence in Molecular Cell Science, Chinese Academy of Sciences, Shanghai 200031, China; University of Chinese Academy of Sciences, Beijing 100049, China. Electronic address: [email protected].
    3
    State Key Laboratory of Molecular Biology, Shanghai Institute of Biochemistry and Cell Biology, Center for Excellence in Molecular Cell Science, Chinese Academy of Sciences, Shanghai 200031, China. Electronic address: [email protected].
    4
    Key Laboratory of Medical Molecular Virology (MOE & MOH), School of Basic Medical Sciences, Fudan University, Shanghai, China 200032. Electronic address: [email protected].
    5
    State Key Laboratory of Molecular Biology, Shanghai Institute of Biochemistry and Cell Biology, Center for Excellence in Molecular Cell Science, Chinese Academy of Sciences, Shanghai 200031, China. Electronic address: [email protected].
    6
    State Key Laboratory of Proteomics, Beijing Proteome Research Center, National Center for Protein Sciences (Beijing), Research Unit of Proteomics & Research and Development of New Drug of Chinese Academy of Medical Sciences, Beijing Institute of Lifeomics, Beijing, China 102206. Electronic address: [email protected].
    7
    Medical College, Guizhou University, Guiyang, Guizhou, China 550025. Electronic address: [email protected].
    8
    Department of Infectious Diseases, Huashan Hospital and Key Laboratory of Medical Molecular Virology (MOH & MOE), Shanghai Medical College, Fudan University, 12 Wulumuqi Zhong Road, Shanghai, China, 200040. Electronic address: [email protected].
    9
    Research Unit Analytical Pathology, Helmholtz Zentrum München, Neuherberg, Germany. Electronic address: [email protected].
    10
    Research Unit Analytical Pathology, Helmholtz Zentrum München, Neuherberg, Germany. Electronic address: [email protected].
    11
    State Key Laboratory of Proteomics, Beijing Proteome Research Center, National Center for Protein Sciences (Beijing), Research Unit of Proteomics & Research and Development of New Drug of Chinese Academy of Medical Sciences, Beijing Institute of Lifeomics, Beijing, China 102206. Electronic address: [email protected].
    12
    Department of Infectious Diseases, Huashan Hospital and Key Laboratory of Medical Molecular Virology (MOH & MOE), Shanghai Medical College, Fudan University, 12 Wulumuqi Zhong Road, Shanghai, China, 200040. Electronic address: [email protected].
    13
    Key Laboratory of Medical Molecular Virology (MOE & MOH), School of Basic Medical Sciences, Fudan University, Shanghai, China 200032. Electronic address: [email protected].
    14
    State Key Laboratory of Molecular Biology, Shanghai Institute of Biochemistry and Cell Biology, Center for Excellence in Molecular Cell Science, Chinese Academy of Sciences, Shanghai 200031, China; University of Chinese Academy of Sciences, Beijing 100049, China; Cancer Center, Shanghai Tenth People's Hospital, School of Medicine, Tongji University, Shanghai 200031, China; School of Life Science, Hangzhou Institute for Advance Study, University of Chinese Academy of Sciences, Hangzhou 310024, China. Electronic address: [email protected].

    PMID: 33621634 DOI: 10.1016/j.jhep.2021.02.009

Abstract

Background and aims: Hepatitis B Virus remains to be yet unresolved global threat to human health. It remains incompletely understood how HBV self-restricts in host during most adulthood infections, and multi-omics analyses were performed to systematically interrogate into HBV-host interaction and the life cycle of HBV.

Methods: RNA-sequencing and ribosome profiling were conducted with cell-based models for HBV replication and gene expression. The novel translational events or products hereby detected were then characterized, and functionally assessed in both cell and mouse models. Moreover, quasi-species analyses of HBV subpopulations were conducted with patients at immune tolerance or activation phases, using next- or third-generation sequencing.

Results: We identified EnhI-SL (Enhancer I-stem loop) as a new cis element in HBV genome, and the mutations disrupting EnhI-SL were found to elevate viral polymerase expression. Furthermore, while re-discovering HpZ/P', a previously under-explored isoform of HBV polymerase, we also identified HBxZ as a novel short isoform of HBX and confirmed their existence and functionally characterized them as potent suppressors for HBV gene expression or genome replication. Mechanistically, HpZ/P' was found to repress HBV gene expression partially through interacting with, and sequestering SUPV3L1. The abundances of the HBV mutants either deficient of HpZ/P' or disrupted in EnhI-SL seemed to be diminished upon the activation of host immune system. Finally, SRSF2, a HBV-down-regulated host protein in RNA spliceosome, was found to promote the splicing of viral pre-genomic RNA and HpZ/P' biogenesis.

Conclusion: This study has identified multiple viral self-restricting mechanisms in HBV-host interaction. Particularly, SRSF2-HpZ/P' appeared to constitute another negative feedback mechanism in controlling HBV life-cycle. Targeting host splicing machinery might thus represent a yet under-explored strategy to intervene into HBV-host interaction.

Keywords: EnhI-SL; HBV; HBxZ; HpZ/P’; Ribosome-profiling; Translatomic.

Copyright © 2021 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.

StephenW

跨抗原譜分析揭示了在HBV感染過程中新型的自我限制性病毒-宿主相互作用
袁士林1，廖光宏2，張夢寬3，朱元飛4，王坤5，肖威迪6，蔡偉佳7，閔惠東8，孫娜9，阿克塞爾·沃爾奇10，徐平11，張繼明12，鄧強13，容桂14
隸屬關係
隸屬關係

    1個
    中國科學院上海生物化學與細胞生物學研究所，分子生物學國家重點實驗室，上海200031；中國科學院大學，北京100049電子地址：[email protected]。
    2
    中國科學院上海生物化學與細胞生物學研究所，分子生物學國家重點實驗室，上海200031；中國科學院大學，北京100049電子地址：[email protected]。
    3
    中國科學院上海生物化學與細胞生物學研究所，分子生物學國家重點實驗室，上海200031電子地址：[email protected]。
    4
    復旦大學基礎醫學院，醫學分子病毒學重點實驗室（MOE＆MOH），上海200032。電子地址：[email protected]。
    5
    中國科學院上海生物化學與細胞生物學研究所，分子生物學國家重點實驗室，上海200031電子地址：[email protected]。
    6
    蛋白質組學國家重點實驗室，北京蛋白質組學研究中心，國家蛋白質科學中心（北京），蛋白質組學研究組和中國醫學科學院新藥研究與開發，北京生命科學研究院，北京102206 ：[email protected]。
    7
    貴州大學醫學院，貴州貴陽550025。電子地址：[email protected]。
    8
    復旦大學上海醫學院華山醫院感染科和醫學分子病毒學重點實驗室，上海市上海烏魯木齊中路12號，200040。電子地址：[email protected]。
    9
    分析病理學研究單位，德國紐黑堡的Helmholtz ZentrumMünchen。電子地址：[email protected]。
    10
    分析病理學研究單位，德國紐黑堡的Helmholtz ZentrumMünchen。電子地址：[email protected]。
    11
    蛋白質組學國家重點實驗室，北京蛋白質組學研究中心，國家蛋白質科學中心（北京），蛋白質組學研究組和中國醫學科學院新藥研究與開發，北京生命科學研究院，北京102206 ：[email protected]。
    12
    復旦大學上海醫學院華山醫院感染科和醫學分子病毒學重點實驗室，上海烏魯木齊中路12號，200040，電子地址：[email protected]。
    13
    復旦大學基礎醫學院，醫學分子病毒學重點實驗室（MOE＆MOH），上海200032。電子地址：[email protected]。
    14
    中國科學院上海生物化學與細胞生物學研究所，分子生物學國家重點實驗室，上海200031；中國科學院大學，北京100049；同濟大學醫學院附屬第十人民醫院腫瘤中心，上海200031；中國科學院大學杭州先進研究學院生命科學學院，杭州310024電子地址：[email protected]。

    PMID：33621634 DOI：10.1016 / j.jhep.2021.02.009

抽象的

背景和目的：乙肝病毒仍然是尚未解決的全球人類健康威脅。仍然不完全了解在大多數成年期感染期間宿主體內HBV如何自我限制，並且進行了多組學分析以系統地詢問HBV-宿主相互作用和HBV的生命週期。

方法：使用基於細胞的HBV複製和基因表達模型對RNA測序和核醣體分析。然後表徵由此檢測到的新的翻譯事件或產物，並在細胞和小鼠模型中對其功能進行評估。此外，使用下一代或第三代測序方法對處於免疫耐受或激活階段的患者進行了HBV亞群的準種分析。
結果：我們鑑定出EnhI-SL（增強子I-莖環）是HBV基因組中的新順式元件，並且發現破壞EnhI-SL的突變可提高病毒聚合酶的表達。此外，在重新發現HpZ / P'（先前未充分探索的HBV聚合酶同工型）的同時，我們還鑑定了HBxZ為HBX的新型短同工型，並確認了它們的存在，並在功能上將其表徵為HBV基因表達或基因組複製的有效抑製劑。 。從機理上講，發現HpZ / P'通過與SUPV3L1相互作用和隔離，部分抑制HBV基因的表達。缺乏HpZ / P'或被EnhI-SL破壞的HBV突變體的豐度在宿主免疫系統激活後似乎減少了。最後，發現SRSF2是RNA剪接體中HBV下調的宿主蛋白，可促進病毒前基因組RNA的剪接和HpZ / P'生物合成。

結論：本研究確定了乙型肝炎病毒-宿主相互作用中的多種病毒自限機制。特別是，SRSF2-HpZ / P'似乎構成了控制HBV生命週期的另一種負反饋機制。因此，針對宿主的剪接機制可能代表了尚未深入研究的干預HBV-宿主相互作用的策略。

關鍵字：EnhI-SL；乙肝病毒HBxZ; HpZ / P’；核醣體圖譜；跨學科的。

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