Smidt M P, Wijnholds J, Snippe L, van Keulen G, Ab G
Laboratory of Biochemistry, University of Groningen, The Netherlands.
Biochim Biophys Acta. 1994 Sep 13;1219(1):115-20. doi: 10.1016/0167-4781(94)90253-4.
Activation of the very low density apolipoprotein II (apoVLDL II) gene in chicken liver by estrogen results in the binding of a variety of nuclear proteins including members of the steroid receptor superfamily and the bZip superfamily to the immediate 5' flanking region. In the present study, we have identified a bZip protein from chicken liver as one of the potential binding activities. Its cognate cDNA was cloned from an expression library using a recognition site DNA probe corresponding to part of the apoVLDL II promoter region. By footprinting and gel shift analysis with the recombinant protein from a prokaryotic expression system we have established that the protein binds to at least three different sites in the apoVLDLII promoter region. One of these sites partially overlaps with the major estrogen response element of the gene. Despite the proximity of their binding sites, the estrogen receptor and the bZip protein can bind simultaneously to the very region. Possible implications of this intimate arrangement of binding sites for the activation of the apoVLDL II promoter are discussed.
雌激素激活鸡肝脏中的极低密度载脂蛋白II(apoVLDL II)基因,导致多种核蛋白与紧邻的5'侧翼区域结合,这些核蛋白包括类固醇受体超家族和bZip超家族的成员。在本研究中,我们已鉴定出一种来自鸡肝脏的bZip蛋白是潜在的结合活性蛋白之一。使用与apoVLDL II启动子区域部分对应的识别位点DNA探针,从表达文库中克隆了其同源cDNA。通过用原核表达系统的重组蛋白进行足迹分析和凝胶迁移分析,我们确定该蛋白与apoVLDLII启动子区域中至少三个不同位点结合。这些位点之一与该基因的主要雌激素反应元件部分重叠。尽管它们的结合位点相邻,但雌激素受体和bZip蛋白可以同时结合到同一区域。讨论了结合位点的这种紧密排列对apoVLDL II启动子激活的可能影响。