Johnson P F, Landschulz W H, Graves B J, McKnight S L
Department of Embryology, Carnegie Institution of Washington, Baltimore 21210.
Genes Dev. 1987 Apr;1(2):133-46. doi: 10.1101/gad.1.2.133.
This report describes the identification and purification of a nuclear protein from rat liver that binds selectively to DNA sequences associated with several animal virus enhancers. The binding activity was tracked by direct DNase I footprinting through four steps of biochemical fractionation. These procedures led to the identification of a polypeptide species exhibiting an apparent molecular weight of 20 kD that accounts for enhancer binding activity. DNase I and dimethyl sulfate footprinting assays were used to examine the manner in which the purified protein binds to enhancer elements associated with SV40, murine sarcoma virus, and polyoma virus. The results of these assays indicate that the initial interaction established between the 20-kD protein and each viral enhancer occurs via a common DNA sequence known as the enhancer core homology.
本报告描述了从大鼠肝脏中鉴定和纯化一种核蛋白的过程,该蛋白能选择性地结合与多种动物病毒增强子相关的DNA序列。通过直接的DNase I足迹法在四个生化分级分离步骤中追踪结合活性。这些步骤导致鉴定出一种表观分子量为20 kD的多肽,它具有增强子结合活性。使用DNase I和硫酸二甲酯足迹法检测纯化蛋白与SV40、鼠肉瘤病毒和多瘤病毒相关增强子元件的结合方式。这些检测结果表明,20-kD蛋白与每种病毒增强子之间最初建立的相互作用是通过一种称为增强子核心同源性的共同DNA序列发生的。