核糖体蛋白S6激酶2(RSK2)在HBx增强HBV转录复制中的作用和机理研究

HBx plays an important role in the argumenting of HBV transcription and replication through transactivating HBV promotor. Our previous study demonstrated that HBx upregulating the expression of RSK2 and the inhibition of HBV replication by knocking down RSK2. Therefore, the effect and mechanism of RSK2 in HBx argumenting HBV transcription and replication is important. .Based on HBV replication model in vivo and vitro, we intend to clarify the effect of RSK2 in HBx argumenting HBV transcription and replication by increasing or decreasing the expression of RSK2. We also aim to clarify whether HBx regulates the expression of RSK2 by upstream MAPK pathway. Secondly, the downstream transcription factors which regulated by RSK2 are screened. Furtherly, we intend to elucidate whether HBx affects the binding of the transcription factor to the HBV promoter by RSK2, which can activate the promoter activity and enhance the transcription and replication of HBV. . To clarify the roles host factors in argumenting HBV transcription and replication would help to elucidate the HBV pathogenicity mechanism. It may provide new targets for the development of anti HBV drugs

乙肝病毒X蛋白（HBx）通过反式激活HBV启动子增强HBV转录与复制。我们的研究显示：HBx显著上调RSK2的表达，而RSK2的表达对HBV的复制有影响。HBx是否通过上调RSK2来实现其增强HBV转录复制的作用及机制有待阐明。.本项目拟利用HBV体外细胞及体内小鼠复制模型，并通过调节RSK2的表达水平，明确其在HBx增强HBV RNA转录、HBV DNA复制中的确切作用；利用通路抑制剂，探明HBx上调RSK2表达的信号通路；筛选RSK2调控的下游转录因子，利用HBV启动子报告基因体系、凝胶迁移实验及染色质免疫共沉淀技术，进一步阐明HBx是否通过RSK2调控的转录因子结合HBV启动子，激活启动子活性，从而增强HBV转录与复制。.本项目的实施将有助于阐明HBx通过宿主蛋白增强HBV转录与复制的分子机制，加深对HBV致病机制的理解，可能为HBV药物研发提供新的靶点。

乙肝病毒X蛋白（HBx）通过反式激活HBV启动子增强HBV转录与复制。本项目前期研究显示：HB x显著上调RSK2的表达，而RSK2的表达对HBV的复制有影响。本项目拟阐明HBx是否通过上调RSK2来实现其增强HBV转录复制的作用及机制。本项目利用HBV体外细胞复制模型，并通过调节RSK2的表达水平，明确其在HBx增强HBV DNA复制中的确切作用。利用通路抑制剂，探明HBx上调RSK2表达的信号通路；利用生物信息学预测并进一步验证HBx调控RSK2表达的关键转录因子，结合双荧光素酶报告基因体系，证实HBx通过该关键转录因子调控RSK2的启动子活性；最后利用Co-IP明确HBx与RSK2之间的相互作用。