Miller Myrna M, Jarosinski Keith W, Schat Karel A
Unit of Avian Health, Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853, USA.
J Gen Virol. 2008 Dec;89(Pt 12):2998-3003. doi: 10.1099/vir.0.2008/003103-0.
Expression of enhanced green fluorescent protein (EGFP) under control of the promoter-enhancer of chicken infectious anemia virus (CAV) is increased in an oestrogen receptor-enhanced cell line when treated with oestrogen and the promoter-enhancer binds unidentified proteins that recognize a consensus oestrogen response element (ERE). Co-transfection assays with the CAV promoter and the nuclear receptor chicken ovalbumin upstream promoter transcription factor 1 (COUP-TF1) showed that expression of EGFP was decreased by 50 to 60 % in DF-1 and LMH cells. The CAV promoter that included sequences at and downstream of the transcription start point had less expression than a short promoter construct. Mutation of a putative E box at this site restored expression levels. Electromobility shift assays showed that the transcription regulator delta-EF1 (deltaEF1) binds to this E box region. These findings indicate that the CAV promoter activity can be affected directly or indirectly by COUP-TF1 and deltaEF1.
在雌激素受体增强的细胞系中,当用雌激素处理时,受鸡传染性贫血病毒(CAV)启动子 - 增强子控制的增强型绿色荧光蛋白(EGFP)的表达增加,并且该启动子 - 增强子与识别共有雌激素反应元件(ERE)的未鉴定蛋白质结合。用CAV启动子与核受体鸡卵清蛋白上游启动子转录因子1(COUP - TF1)进行的共转染试验表明,在DF - 1和LMH细胞中,EGFP的表达降低了50%至60%。包含转录起始点及其下游序列的CAV启动子的表达低于短启动子构建体。该位点推定的E盒发生突变后恢复了表达水平。电泳迁移率变动分析表明,转录调节因子δ - EF1(deltaEF1)与该E盒区域结合。这些发现表明,CAV启动子活性可直接或间接受到COUP - TF1和deltaEF1的影响。