HBV X蛋白结合共同转录因子ⅡB的研究

• 作 者: 杨益大 马亦林 郑临 村上清史
• 机 构: 浙江大学附属第一医院,日本金泽大学肿瘤研究所
• 出 处: 中华传染病杂志
• 发表日期: 1999年03期
• 关键词: 蛋白结合 乙型肝炎病毒 HBV X蛋白 转录因子 病毒基因 免疫沉淀试验 复制水平 融合蛋白 亚单位

目的研究乙型肝炎病毒Ｘ蛋白（ＨＢＶＸ蛋白）能否结合共同转录因子ⅡＢ（ＴＦⅡＢ）。方法应用体外蛋白与蛋白结合试验（ＰｕｌＤｏｗｎ试验）和ＦａｒＷｅｓｔｅｒｎＢｌｏｔ试验，以及细胞内联合免疫沉淀试验。结果ＨＢＶＸ蛋白能直接与普通ＴＦⅡＢ结合。ＨＢＶＸ的Ｘ１９９（５１－９９）是结合ＴＦⅡＢ的最小基团。研究还证实土拨鼠肝炎病毒Ｘ蛋白（ＷＨＸ）同样能与ＴＦⅡＢ结合。细胞内联合免疫沉淀试验更进一步证实了体外蛋白与蛋白结合实验的结果。结论嗜肝ＤＮＡ病毒Ｘ蛋白与ＴＦⅡＢ结合是其发挥反式转录激活作用的机制之一。

Objective To investigate whether hepatitis B virus X protein (HBVX protein) can bind to common transcription factor ⅡB (TFⅡB). Methods The protein-protein binding assay (PulDown test) and FarWesternBlot assay were performed in vitro and intracellular immunoprecipitation assay was performed. The results of HBVX protein can be directly associated with ordinary TF Ⅱ B binding. X199 (51-99) of HBVX is the smallest group that binds to TFIIB. The study also confirmed that woodchuck hepatitis virus X protein (WHX) also binds to TFIIB. Intracellular immunoprecipitation assay further confirmed the results of in vitro protein-protein binding experiments. Conclusion The combination of hepadnavirus X protein with TFⅡB is one of the mechanisms of its transcriptional activation.