Rabek J P, Papaconstantinou J
Department of Human Biological Chemistry and Genetics, University of Texas Medical Branch, Galveston, Texas, 77555-0643, USA.
Biochem Biophys Res Commun. 1999 Feb 24;255(3):608-13. doi: 10.1006/bbrc.1999.0240.
Exchange in binding of transcription factors C/EBPalpha and C/EBPbeta at a regulatory site in the alpha1-acid glycoprotein (AGP) promoter, termed the acute phase response element (APRE), has been correlated with bacterial lipopolysaccharide (LPS) mediated induction. The APRE contains overlapping recognition sequences for C/EBP's and glucocorticoid receptor (GR). Electrophortetic mobility shift assays show that this site can bind both GR and C/EBP. However, using liver nuclear extract, which contains GR binding activity, only C/EBP binds to the APRE. Binding interference methods, using dimethyl sulfate and potassium permanganate modification of specific bases, detected interference only with modification of bases that are in the region of the C/EBP binding site that do not overlap with the GRE sequence. There are no significant differences between the interference patterns of control and LPS treated liver nuclear extracts, suggesting that the region of close contact between protein and DNA is similar for C/EBPalpha (untreated) and C/EBPbeta (treated).
转录因子C/EBPα和C/EBPβ在α1-酸性糖蛋白(AGP)启动子的一个调控位点(称为急性期反应元件,APRE)上的结合交换,已与细菌脂多糖(LPS)介导的诱导相关。APRE包含C/EBP和糖皮质激素受体(GR)的重叠识别序列。电泳迁移率变动分析表明,该位点可同时结合GR和C/EBP。然而,使用含有GR结合活性的肝核提取物时,只有C/EBP与APRE结合。利用硫酸二甲酯和高锰酸钾对特定碱基进行修饰的结合干扰方法,仅检测到对C/EBP结合位点区域中不与GRE序列重叠的碱基修饰的干扰。对照和LPS处理的肝核提取物的干扰模式之间没有显著差异,这表明C/EBPα(未处理)和C/EBPβ(处理)与蛋白质和DNA紧密接触的区域相似。