Kovesdi I, Reichel R, Nevins J R
Cell. 1986 Apr 25;45(2):219-28. doi: 10.1016/0092-8674(86)90386-7.
We have employed a gel assay to detect a transcription factor in nuclear extracts of adenovirus-infected cells that interacts with the adenovirus E2 promoter, an E1A inducible promoter. Binding of the factor to the promoter protected sequences between -33 and -74 from DNAase cleavage in a footprint assay. This region was also protected from exonuclease III digestion. These sequences coincide with those previously shown to be essential for promoter activity and E1A stimulation. The factor could be detected in extracts of uninfected cells, although at greatly reduced levels. The increased level of factor in infected cells required a functional E1A gene. These results suggest that the E2 binding activity is a cellular transcriptional factor, the concentration or binding activity of which increases as a result of the action of the E1A gene product.
我们采用凝胶分析法来检测腺病毒感染细胞的核提取物中的一种转录因子,该因子可与腺病毒E2启动子相互作用,E2启动子是一种E1A诱导型启动子。在足迹分析中,该因子与启动子的结合保护了-33至-74之间的序列不被DNA酶切割。该区域也免受核酸外切酶III的消化。这些序列与先前显示对启动子活性和E1A刺激至关重要的序列一致。尽管未感染细胞提取物中的该因子水平大幅降低,但仍可检测到。感染细胞中该因子水平的升高需要一个功能性的E1A基因。这些结果表明,E2结合活性是一种细胞转录因子,其浓度或结合活性因E1A基因产物的作用而增加。