乙型肝炎病毒核心蛋白核相互作用组确定SRSF10是限制HBV RNA产

StephenW

Hepatitis B virus Core protein nuclear interactome identifies SRSF10 as a host RNA-binding protein restricting HBV RNA production
Hélène Chabrolles  1 , Héloïse Auclair  1 , Serena Vegna  1 , Thomas Lahlali  1 , Caroline Pons  1 , Maud Michelet  1 , Yohann Couté  2 , Lucid Belmudes  2 , Gilliane Chadeuf  3 , Yujin Kim  1 , Ariel Di Bernardo  1 , Pascal Jalaguier  1 , François-Loïc Cosset  4 , Floriane Fusil  4 , Michel Rivoire  5 , Lee D Arnold  6 , Uri Lopatin  7 , Christophe Combet  1 , Fabien Zoulim  1 , David Grierson  8 , Benoit Chabot  9 , Julie Lucifora  1 , David Durantel  1 , Anna Salvetti  1
Affiliations
Affiliations

    1
    INSERM, U1052, Cancer Research Center of Lyon (CRCL), Université de Lyon (UCBL1), CNRS UMR5286, Centre Léon Bérard, Lyon, France.
    2
    Univ. Grenoble Alpes, CEA, INSERM, IRIG, BGE, Grenoble, France.
    3
    INSERM U1087, Institut du Thorax, Université de Nantes, CNRS UMR6291, Nantes, France.
    4
    INSERM, U1111, International Center for Infectiology Research (CIRI), Université de Lyon (UCBL1), CNRS, UMR5308, Ecole Normale Supérieure de Lyon, Lyon, France.
    5
    INSERM U1032, Centre Léon Bérard (CLB), Lyon, France.
    6
    DiscoverElucidations, LLC, Rancho Santa Fe, California, United States of America.
    7
    Assembly Biosciences, San Francisco, California, United States of America.
    8
    Faculty of Pharmaceutical Sciences, University of British Columbia, Vancouver, British Columbia, Canada.
    9
    Department of Microbiology and Infectious Diseases, Faculty of Medicine and Health Sciences, Université de Sherbrooke, Sherbrooke, Quebec, Canada.

    PMID: 33180834 DOI: 10.1371/journal.ppat.1008593

Abstract

Despite the existence of a preventive vaccine, chronic infection with Hepatitis B virus (HBV) affects more than 250 million people and represents a major global cause of hepatocellular carcinoma (HCC) worldwide. Current clinical treatments, in most of cases, do not eliminate viral genome that persists as a DNA episome in the nucleus of hepatocytes and constitutes a stable template for the continuous expression of viral genes. Several studies suggest that, among viral factors, the HBV core protein (HBc), well-known for its structural role in the cytoplasm, could have critical regulatory functions in the nucleus of infected hepatocytes. To elucidate these functions, we performed a proteomic analysis of HBc-interacting host-factors in the nucleus of differentiated HepaRG, a surrogate model of human hepatocytes. The HBc interactome was found to consist primarily of RNA-binding proteins (RBPs), which are involved in various aspects of mRNA metabolism. Among them, we focused our studies on SRSF10, a RBP that was previously shown to regulate alternative splicing (AS) in a phosphorylation-dependent manner and to control stress and DNA damage responses, as well as viral replication. Functional studies combining SRSF10 knockdown and a pharmacological inhibitor of SRSF10 phosphorylation (1C8) showed that SRSF10 behaves as a restriction factor that regulates HBV RNAs levels and that its dephosphorylated form is likely responsible for the anti-viral effect. Surprisingly, neither SRSF10 knock-down nor 1C8 treatment modified the splicing of HBV RNAs but rather modulated the level of nascent HBV RNA. Altogether, our work suggests that in the nucleus of infected cells HBc interacts with multiple RBPs that regulate viral RNA metabolism. Our identification of SRSF10 as a new anti-HBV restriction factor offers new perspectives for the development of new host-targeted antiviral strategies.
Conflict of interest statement

I have read the journal's policy and the authors of this manuscript have the following competing interests: Uri Lopatin is an advisor to and shareholder of Assembly Biosciences. Lee Arnold was an employee of Assembly Biosciences.

StephenW

乙型肝炎病毒核心蛋白核相互作用组确定SRSF10是限制HBV RNA产生的宿主RNA结合蛋白
HélèneChabrolles 1，HéloïseAuclair 1，Serena Vegna 1，Thomas Lahlali 1，Caroline Pons 1，Maud Michelet 1，YohannCouté2，Lucid Belmudes 2，Gilliane Chadeuf 3，Yujin Kim 1，Ariel Di Bernardo 1，Pascal Jalaguier 1，François -LoïcCosset 4，Floriane Fusil 4，Michel Rivoire 5，Lee D Arnold 6，Uri Lopatin 7，Christophe Combet 1，Fabien Zoulim 1，David Grierson 8，Benoit Chabot 9，Julie Lucifora 1，David Durantel 1，Anna Salvetti 1
隶属关系
隶属关系

    1个
    INSERM，U1052，里昂大学癌症研究中心（CRCL），里昂大学（UCBL1），CNRS UMR5286，里昂贝拉德中心，法国。
    2
    大学格勒诺布尔Alpes，CEA，INSERM，IRIG，BGE，格勒诺布尔，法国。
    3
    法国南特CNRS UMR6291，南特大学Thorax研究所INSERM U1087。
    4
    INSERM，U1111，里昂大学国际感染研究中心（CIRI），UCRS，CNRS，UMR5308，里昂高等师范学院，法国里昂。
    5
    INSERM U1032，法国里昂市中心贝拉德（CLB）。
    6
    美国加利福尼亚州圣达菲牧场的DiscoverElucidations，LLC。
    7
    Assembly Biosciences，美国加利福尼亚州旧金山。
    8
    加拿大不列颠哥伦比亚省温哥华市不列颠哥伦比亚大学药学院。
    9
    加拿大魁北克，舍布鲁克，舍布鲁克大学，医学与健康科学学院，微生物学和传染病系。

    PMID：33180834 DOI：10.1371 / journal.ppat.1008593

抽象

尽管存在预防性疫苗，但乙型肝炎病毒（HBV）的慢性感染仍影响着超过2.5亿人，并代表了全球范围内肝细胞癌（HCC）的主要全球原因。在大多数情况下，当前的临床治疗并未消除病毒基因组，该病毒基因组以DNA附加体的形式存在于肝细胞核中，并构成了病毒基因连续表达的稳定模板。几项研究表明，在病毒因素中，以其在细胞质中的结构作用而闻名的HBV核心蛋白（HBc）在受感染的肝细胞核中可能具有关键的调节功能。为了阐明这些功能，我们在分化的HepaRG（人类肝细胞的替代模型）的细胞核中进行了与HBc相互作用的宿主因子的蛋白质组学分析。发现HBc相互作用组主要由RNA结合蛋白（RBP）组成，这些蛋白与mRNA代谢的各个方面有关。其中，我们的研究重点是SRSF10，这是一种RBP，以​​前显示它可以以磷酸化依赖性方式调节替代剪接（AS），并控制应激和DNA损伤反应以及病毒复制。结合SRSF10敲低和SRSF10磷酸化的药理抑制剂的功能研究表明，SRSF10充当调节HBV RNA水平的限制因子，其去磷酸化形式可能是抗病毒作用的原因。出人意料的是，SRSF10敲除或1C8处理均未改变HBV RNA的剪接，而是调节了新生HBV RNA的水平。总之，我们的研究表明，在被感染细胞的核中，HBc与调节病毒RNA代谢的多个RBP相互作用。我们将SRSF10鉴定为一种新的抗HBV限制因子，为开发新的针对宿主的抗病毒策略提供了新的视角。
利益冲突声明

我已经阅读了该杂志的政策，该手稿的作者有以下竞争利益：Uri Lopatin是Assembly Biosciences的顾问和股东。 Lee Arnold是Assembly Biosciences的一名员工。

StephenW

https://journals.plos.org/plospa ... &type=printable