Notch signaling facilitates hepatitis B virus covalently closed circular DNA transcription via cAMP response element-binding protein with E3 ubiquitin ligase-modulation
Notch signaling facilitates hepatitis B virus covalently closed circular DNA transcription via cAMP response element-binding protein with E3 ubiquitin ligase-modulation
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DOI：
10.1038/s41598-018-38139-5
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发表时间：
2019-02
影响因子：
4.6
通讯作者：
Zijing Wang;K. Kawaguchi;M. Honda;S. Hashimoto;Takayoshi Shirasaki;Hikari Okada;Noriaki Orita;Tetsuro Shimakami;T. Yamashita;Y. Sakai;E. Mizukoshi;S. Murakami;S. Kaneko
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文献类型：
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作者：
Zijing Wang;K. Kawaguchi;M. Honda;S. Hashimoto;Takayoshi Shirasaki;Hikari Okada;Noriaki Orita;Tetsuro Shimakami;T. Yamashita;Y. Sakai;E. Mizukoshi;S. Murakami;S. Kaneko
NOTCH1受E3泛素连接酶调控，Notch胞内区的蛋白酶体降解影响靶基因的转录。CAMP反应元件结合蛋白(CREB)介导乙肝病毒(乙肝病毒)共价闭合环状DNA(CccDNA)的转录。我们评估了HBVcccDNA与Notch信号活性的关系。用Jagged1shRNA和γ分泌酶抑制剂处理乙肝病毒复制细胞，检测HBVccDNA水平和相关基因的表达。这些因素在手术切除的临床样本中的影响也被评估。缺口抑制抑制了HBVcccDNA和CREB相关的表达，但增加了瘙痒和麻木水平。蛋白酶体抑制剂增强HBVccDNA，恢复Notch和CREB的表达，并抑制瘙痒和Numb功能。在瘙痒和麻木阻断后，甚至在腺苷环化酶激活剂Forsklin治疗后，仍可观察到HBVccDNA的升高；蛋白激酶A(PKA)抑制剂则有相反的作用。临床肝组织中HBVDNA阳性细胞有缺口激活和E3连接酶失活现象。综上所述，这些发现表明，Notch信号活性通过CREB促进HBVcccDNA的转录，从而触发下游的PKA-磷酸化-CREB级联反应，并受Notch胞内域E3泛素连接酶的调节。
Notch1 is regulated by E3 ubiquitin ligases, with proteasomal degradation of the Notch intracellular domain affecting the transcription of target genes. cAMP response element-binding protein (CREB) mediates the transcription of hepatitis B virus (HBV) covalently closed circular DNA (cccDNA). We assessed the relationship between HBV cccDNA and Notch signaling activities. HBV cccDNA levels and relative gene expression were evaluated in HBV-replicating cells treated with Jagged1 shRNA and a γ-secretase inhibitor. The effects of these factors in surgically resected clinical samples were also assessed. Notch inhibition suppressed HBV cccDNA and CREB-related expression but increased ITCH and NUMB levels. Proteasome inhibitor augmented HBV cccDNA, restored Notch and CREB expression, and inhibited ITCH and NUMB function. Increased HBV cccDNA was observed after ITCH and NUMB blockage, even after treatment with the adenylate cyclase activator forskolin; protein kinase A (PKA) inhibitor had the opposite effect. Notch activation and E3 ligase inactivation were observed in HBV-positive cells in clinical liver tissue. Collectively, these findings reveal that Notch signaling activity facilitates HBV cccDNA transcription via CREB to trigger the downstream PKA-phospho-CREB cascade and is regulated by E3 ubiquitin ligase-modulation of the Notch intracellular domain.