目的:筛选参与APOBEC3B羧基端脱氨酶及抗乙肝病毒(hepatitis B virus,HBV)活性的重要区域和关键位点。方法:定点突变构建羧基端活性中心附近的3个螺旋区段(α2、α3、α4)的缺失突变体和α4螺旋区内(D316、K320、E321、R327、D328)位点...目的:筛选参与APOBEC3B羧基端脱氨酶及抗乙肝病毒(hepatitis B virus,HBV)活性的重要区域和关键位点。方法:定点突变构建羧基端活性中心附近的3个螺旋区段(α2、α3、α4)的缺失突变体和α4螺旋区内(D316、K320、E321、R327、D328)位点的突变体;Southern blot筛选APOBEC3B抑制HBV复制的重要区段和关键位点;不同DNA变性PCR(differential DNA denaturation PCR,3D-PCR)和克隆测序进行验证。结果:APOBEC3B羧基端脱氨酶活性中心周围的α3和α4螺旋区缺失后,其抗病毒活性消失;D316位点突变后APOBEC3B的脱氨酶活性和抗病毒活性消失。结论:APOBEC3B羧基端的D316位点是APOBEC3B脱氨酶和抗病毒的关键位点。展开更多
APOBEC3 protein families, a DNA cytidine deaminase, were up-regulated in multiple tumors. However, the relationship between Hepatocellular carcinoma(HCC) and APOBEC3B(A3B) remains unknown. It has been confirmed that i...APOBEC3 protein families, a DNA cytidine deaminase, were up-regulated in multiple tumors. However, the relationship between Hepatocellular carcinoma(HCC) and APOBEC3B(A3B) remains unknown. It has been confirmed that interleukin-6(IL-6)has significant impacts on oncogenesis of HCC. Here, we reported that the expression of IL-6 was substantially up-regulated by A3 B in HepG2 cells. A3 B induced IL-6 expression through relocating HuR to enhance the IL-6 mRNA stability. Further analysis indicated that IL-6 also increased the expression of A3 B through JAK1/STAT3 signaling pathway, which formed a positive feedback to maintain the continuous expression of A3 B and IL-6, and thereby promoted the prolonged non-resolving inflammation. Collectively, these findings suggest that A3 B is essential for oncogenesis of HCC, and is a potential target for preventive intervention.展开更多
目的研究热休克蛋白70(heat shock protein 70,HSP70)在载脂蛋白B mRNA编辑酶催化多肽样蛋白3B(apolipoprotein B mRNA editing enzyme catalytic polypeptide-like 3B,APOBEC3B)抑制乙型肝炎病毒(hepatitis B virus,HBV)复制过程中的...目的研究热休克蛋白70(heat shock protein 70,HSP70)在载脂蛋白B mRNA编辑酶催化多肽样蛋白3B(apolipoprotein B mRNA editing enzyme catalytic polypeptide-like 3B,APOBEC3B)抑制乙型肝炎病毒(hepatitis B virus,HBV)复制过程中的作用及机制。方法通过免疫共沉淀(co-immunopreciptation,Co-IP)和GST pull-down试验研究HSP70与APOBEC3B的相互作用。干扰HSP70、过表达HSP70或使用HSP70抑制剂VER155008处理肝细胞后,Southern blot或real-time PCR检测APOBEC3B抗病毒作用的改变;差异DNA变性PCR(differential DNA denaturation PCR,3D-PCR)和克隆测序检测APOBEC3B脱氨酶活性的改变。结果HSP70能与APOBEC3B相结合,过表达HSP70能增强APOBEC3B的脱氨酶活性和抗HBV功能。相反,干扰HSP70或使用HSP70抑制剂VER155008后,APOBEC3B脱氨酶活性和抗HBV功能均减弱。结论HSP70通过增强APOBEC3B的脱氨酶活性增强其抗病毒功能。展开更多
文摘目的:筛选参与APOBEC3B羧基端脱氨酶及抗乙肝病毒(hepatitis B virus,HBV)活性的重要区域和关键位点。方法:定点突变构建羧基端活性中心附近的3个螺旋区段(α2、α3、α4)的缺失突变体和α4螺旋区内(D316、K320、E321、R327、D328)位点的突变体;Southern blot筛选APOBEC3B抑制HBV复制的重要区段和关键位点;不同DNA变性PCR(differential DNA denaturation PCR,3D-PCR)和克隆测序进行验证。结果:APOBEC3B羧基端脱氨酶活性中心周围的α3和α4螺旋区缺失后,其抗病毒活性消失;D316位点突变后APOBEC3B的脱氨酶活性和抗病毒活性消失。结论:APOBEC3B羧基端的D316位点是APOBEC3B脱氨酶和抗病毒的关键位点。
基金supported in part by the National Natural Science Foundation of China (91529304, 81502407, 81473230, 81273547, 81403020 and 91129731)the Jiangsu Province Natural Science Foundation of China (BK2014666)the Central College Fund (ZJ14020)
文摘APOBEC3 protein families, a DNA cytidine deaminase, were up-regulated in multiple tumors. However, the relationship between Hepatocellular carcinoma(HCC) and APOBEC3B(A3B) remains unknown. It has been confirmed that interleukin-6(IL-6)has significant impacts on oncogenesis of HCC. Here, we reported that the expression of IL-6 was substantially up-regulated by A3 B in HepG2 cells. A3 B induced IL-6 expression through relocating HuR to enhance the IL-6 mRNA stability. Further analysis indicated that IL-6 also increased the expression of A3 B through JAK1/STAT3 signaling pathway, which formed a positive feedback to maintain the continuous expression of A3 B and IL-6, and thereby promoted the prolonged non-resolving inflammation. Collectively, these findings suggest that A3 B is essential for oncogenesis of HCC, and is a potential target for preventive intervention.
文摘目的研究热休克蛋白70(heat shock protein 70,HSP70)在载脂蛋白B mRNA编辑酶催化多肽样蛋白3B(apolipoprotein B mRNA editing enzyme catalytic polypeptide-like 3B,APOBEC3B)抑制乙型肝炎病毒(hepatitis B virus,HBV)复制过程中的作用及机制。方法通过免疫共沉淀(co-immunopreciptation,Co-IP)和GST pull-down试验研究HSP70与APOBEC3B的相互作用。干扰HSP70、过表达HSP70或使用HSP70抑制剂VER155008处理肝细胞后,Southern blot或real-time PCR检测APOBEC3B抗病毒作用的改变;差异DNA变性PCR(differential DNA denaturation PCR,3D-PCR)和克隆测序检测APOBEC3B脱氨酶活性的改变。结果HSP70能与APOBEC3B相结合,过表达HSP70能增强APOBEC3B的脱氨酶活性和抗HBV功能。相反,干扰HSP70或使用HSP70抑制剂VER155008后,APOBEC3B脱氨酶活性和抗HBV功能均减弱。结论HSP70通过增强APOBEC3B的脱氨酶活性增强其抗病毒功能。
