Hepatitis B virus Core protein nuclear interactome identifies SRSF10 as a host RNA-binding protein restricting HBV RNA production.
Hepatitis B virus Core protein nuclear interactome identifies SRSF10 as a host RNA-binding protein restricting HBV RNA production.
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DOI：
10.1371/journal.ppat.1008593
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发表时间：
2020-11
期刊：
影响因子：
6.7
通讯作者：
Salvetti A
中科院分区：
文献类型：
--
作者：
Chabrolles H;Auclair H;Vegna S;Lahlali T;Pons C;Michelet M;Couté Y;Belmudes L;Chadeuf G;Kim Y;Di Bernardo A;Jalaguier P;Cosset FL;Fusil F;Rivoire M;Arnold LD;Lopatin U;Combet C;Zoulim F;Grierson D;Chabot B;Lucifora J;Durantel D;Salvetti A
尽管存在预防性疫苗，但慢性乙肝病毒感染影响着超过2.5亿人，是全球肝细胞癌的主要原因。目前的临床治疗，在大多数情况下，并没有消除病毒基因组，它以DNA Episome的形式存在于肝细胞核中，并构成病毒基因持续表达的稳定模板。一些研究表明，在病毒因子中，众所周知的乙肝病毒核心蛋白(HBC)在细胞质中的结构作用，可能在受感染的肝细胞的核中具有关键的调节功能。为了阐明这些功能，我们对分化的HepaRG核中与HBC相互作用的宿主因子进行了蛋白质组学分析，分化的HepaRG是人类肝细胞的代理模型。HBC相互作用组主要由RNA结合蛋白(RNAbindingProteins，RBPs)组成，这些蛋白参与了mRNA代谢的各个方面。其中，我们的研究重点是SRSF10，这是一种RBP，以前被证明以磷酸化依赖的方式调节选择性剪接(AS)，控制应激和DNA损伤反应，以及病毒复制。结合SRSF10基因敲除和SRSF10磷酸化的药物抑制物(1C8)的功能研究表明，SRSF10作为一个限制因子调节HBVRNA水平，其去磷酸化形式可能与抗病毒作用有关。令人惊讶的是，SRSF10和1C8治疗都没有改变HBVRNA的剪接，而是调节了新生的HBVRNA的水平。总之，我们的工作表明，在感染细胞的细胞核中，HBC与调节病毒RNA代谢的多个限制性商业惯例相互作用。我们发现SRSF10是一种新的抗乙肝病毒限制因子，为开发新的宿主靶向抗病毒策略提供了新的视角。慢性乙肝病毒感染影响着全球超过2.5亿人，是全球肝癌的主要原因。目前的治疗方法显著减少了患者的病毒血症。然而，病毒很少被清除，而病毒基因组在肝细胞核中的持续存在产生了稳定的病毒RNA来源，从而产生了在免疫逃逸机制和肝病进展中发挥重要作用的蛋白质。仍然需要旨在有效和持久地消除病毒基因表达的治疗方法。在这项研究中，我们确定了乙肝病毒核心蛋白(HBC)的核伙伴，以了解这种负责在细胞质中组装衣壳的结构蛋白如何也能调节病毒基因的表达。HBC相互作用组主要由RNA结合蛋白(RBPs)组成。其中一种限制性商业惯例SRSF10被证明可以限制HBVRNA水平，一种能够改变其磷酸化的药物表现为一种能够减少病毒基因表达的抗病毒化合物。总之，这项研究揭示了HBC的新的调控功能，并为开发旨在防止病毒基因表达的抗病毒策略提供了相关信息。
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. Chronic infection with Hepatitis B virus (HBV) affects more than 250 million of people world-wide and is a major global cause of liver cancer. Current treatments lead to a significant reduction of viremia in patients. However, viral clearance is rarely obtained and the persistence of the HBV genome in the hepatocyte’s nucleus generates a stable source of viral RNAs and subsequently proteins which play important roles in immune escape mechanisms and liver disease progression. Therapies aiming at efficiently and durably eliminating viral gene expression are still required. In this study, we identified the nuclear partners of the HBV Core protein (HBc) to understand how this structural protein, responsible for capsid assembly in the cytoplasm, could also regulate viral gene expression. The HBc interactome was found to consist primarily of RNA-binding proteins (RBPs). One of these RBPs, SRSF10, was demonstrated to restrict HBV RNA levels and a drug, able to alter its phosphorylation, behaved as an antiviral compound capable of reducing viral gene expression. Altogether, this study sheds new light on novel regulatory functions of HBc and provides information relevant for the development of antiviral strategies aiming at preventing viral gene expression.
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