Jose-Estanyol M, Poliard A, Foiret D, Danan J L
Laboratoire d'Enzymologie, Centre National de la Recherche Scientifique, Gif-sur-Yvette, France.
Eur J Biochem. 1989 May 15;181(3):761-6. doi: 10.1111/j.1432-1033.1989.tb14789.x.
Rat liver nuclear extracts were tested for the presence of factors which might be common to the transcriptional regulation of both the albumin and alpha-foetoprotein genes. Gel shift assay showed the formation of three complexes (I, II and III) with the albumin probe. Two of them (I and III) could be displaced by the alpha-foetoprotein promoter. Analysis of nuclear extracts from liver, kidney, spleen and brain and competition experiments using several oligonucleotides covering regions from the albumin and alpha-foetoprotein promoters showed that complex III results from the binding of the ubiquitous nuclear factor 1, while complex II involves a CCAAT-box-binding protein also detected in brain and spleen extracts. Complex I is formed upon binding of a liver-specific factor to a proximal element of the rat albumin promoter. This factor also binds to a similar sequence in the alpha-foetoprotein promoter and is closely related to the hepatocyte nuclear factor 1, as shown by competition experiments using an oligonucleotide covering its target sequence on the beta-fibrinogen promoter. Transfection competition experiments indicated that, in vivo, this factor acts as a positive trans-acting element in the expression of both the rat albumin and alpha-foetoprotein genes.
对大鼠肝脏核提取物进行检测,以寻找可能在白蛋白基因和甲胎蛋白基因转录调控中共同存在的因子。凝胶迁移实验显示,白蛋白探针形成了三种复合物(I、II和III)。其中两种(I和III)可被甲胎蛋白启动子取代。对肝脏、肾脏、脾脏和大脑的核提取物进行分析,并使用覆盖白蛋白和甲胎蛋白启动子区域的几种寡核苷酸进行竞争实验,结果表明复合物III是由普遍存在的核因子1结合形成的,而复合物II涉及一种在大脑和脾脏提取物中也检测到的CCAAT盒结合蛋白。复合物I是由一种肝脏特异性因子与大鼠白蛋白启动子的近端元件结合形成的。通过使用覆盖其在β-纤维蛋白原启动子上靶序列的寡核苷酸进行竞争实验表明,该因子也与甲胎蛋白启动子中的类似序列结合,并且与肝细胞核因子1密切相关。转染竞争实验表明,在体内,该因子在大鼠白蛋白基因和甲胎蛋白基因的表达中作为一种正向反式作用元件发挥作用。