HBV X蛋白结合共同转录因子ⅡB的研究

Study on the binding of HBV X protien to the general transcription factor

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 study the binding of HBV X protein(HBV X) to the general transcription factor B(TF B). Methods PullDown assay in vitro, FarWestern Blot assay and CoImmunoprecipitation assay in vivo were used to detect the binding of HBX protien to TF B. Results It was found that HBX binded TF B directly and HBV X 199(51-99)is the minimal domain responsible for the binding to TF B. Furthermore, the binding of TF BHBX in vitro was confirmed by intracellular coimmunoprecripition assay. It was also found that the woodchuck hepatitis virus X protien (WHX) was to bind TF B. Conclusion These RE results suggest that the interaction between HBX and TF B maybe of the mechanisms which account for its transcription activity.