NUCLEIC ACID FOOTPRINTING OF INTERACTION OF TATA BINDING PROTEIN (TBP) WITH DNA

TBP是核蛋白复合物的组成部分， 三种真核RNA中的每一种的转录起始 聚合酶 的结合 TATA结合蛋白 (TBP)到 特异性启动子序列称为 TATA盒 这是一个关键步骤， 启动由RNA聚合酶II转录的基因的转录。 的核苷酸序列 TATA盒 在自然界中是可变的 启动子发生。 同步辐射X射线核酸的应用 酸和蛋白质足迹对蛋白质-DNA相互作用的影响 通过约束 TATA结合蛋白 (TBP)到 特异性启动子序列。 这种蛋白质是不寻常的，因为它 在结构上引起剧烈的构象变化 的DNA;特别感兴趣的是两个扭结的引入， 通过插入两对苯丙氨酸残基来修饰DNA。 基于其他类型的动力学数据， 这些嵌入事件可以顺序进行，并且它们的 速率可能取决于DNA序列。 滴定实验已经 这表明，准确的同步加速器 该系统可以采集足迹数据。 现在的舞台是 同步辐射足迹动力学的初始化 将严格检验这一假设的实验。

TBP is a component of the nucleo-protein complexes required for the initiation of transcription by each of the three eukaryotic RNA polymerases. The binding of the TATA Binding Protein (TBP) to specific promoter sequences called TATA boxes is a key step in the initiation of transcription of genes transcribed by RNA polymerase II. The nucleotide se-quence of TATA boxes is variable among naturally occurring promoters. The application of synchrotron x-ray nucleic acid and protein footprinting to protein-DNA interactions is being addressed through the binding of the TATA Binding Protein (TBP) to specific promoter se-quences. This protein is unusual in that it induces a dramatic and drastic conformational change in the structure of the DNA; of particular interest is the introduction of two kinks in Hthe DNA by the intercalation of two pairs of phenylalanine residues. HIt has been proposed based on other types of kinetics data, that these Hintercalation events may proceed sequentially and that their rates may Hbe DNA sequence dependent. Titration experiments have been Hsuc-cessfully conducted which demonstrate that accurate synchrotron Hfootprinting data can be acquired for this system. The stage is now Hset for the initiation of synchrotron footprinting ki-netics Hexperiments that will critically test this hypothesis.