Smidt M P, Snippe L, van Keulen G, Ab G
Laboratory of Biochemistry, University of Groningen, The Netherlands.
Eur J Biochem. 1998 Aug 15;256(1):106-11. doi: 10.1046/j.1432-1327.1998.2560106.x.
The vitellogenin-binding protein (VBP) is a member of the proline and acidic-region rich (PAR) family of bZip transcription factors. PAR is located N-terminally to the DNA-binding domain. VBP binds to specific sites within the 300-bp 5'-flanking region of the chicken-liver-specific estrogen-dependent very-low-density apolipoprotein gene (apoVLDL II). One of these binding sites (site D) resembles the albumin site D and is positioned in close proximity of the major estrogen-responsive element. Previous studies showed that VBP can bind simultaneously with the estrogen receptor to the putative complex regulatory element E1D. To investigate whether VBP is involved in apoVLDL II gene expression, we examined its capacity to enhance apoVLDL II transcription and its presence in liver. We show that VBP is capable of enhancing transcription in transfection experiments. However, VBP could not be detected in liver by Western-blots or immuno-electro mobility shift assays (EMSAs) using antibodies against different moieties of the protein. We examined the possible reduction in translation efficiencies due to a small upstream open reading frame in the VBP leader sequence, but did not find any. Although VBP binds to the proximal apoVLDL II promoter region and enhances transcription in co-transfection experiments, the protein is unlikely to be involved in apoVLDL II gene transcription because of its undetectable low level in liver nuclei.
卵黄蛋白原结合蛋白(VBP)是富含脯氨酸和酸性区域(PAR)的bZip转录因子家族成员。PAR位于DNA结合结构域的N端。VBP与鸡肝脏特异性雌激素依赖性极低密度载脂蛋白基因(apoVLDL II)300bp 5'侧翼区域内的特定位点结合。其中一个结合位点(位点D)类似于白蛋白位点D,且位于主要雌激素反应元件附近。先前的研究表明,VBP可与雌激素受体同时结合到假定的复合调控元件E1D上。为了研究VBP是否参与apoVLDL II基因表达,我们检测了其增强apoVLDL II转录的能力及其在肝脏中的存在情况。我们发现VBP在转染实验中能够增强转录。然而,使用针对该蛋白不同部分的抗体,通过蛋白质免疫印迹法或免疫电泳迁移率变动分析(EMSA)在肝脏中均未检测到VBP。我们研究了由于VBP前导序列中的小上游开放阅读框导致翻译效率可能降低的情况,但未发现任何影响。尽管VBP在共转染实验中与apoVLDL II近端启动子区域结合并增强转录,但由于在肝细胞核中检测不到其低水平表达,该蛋白不太可能参与apoVLDL II基因转录。