Constitutive and metal-inducible protein:DNA interactions at the mouse metallothionein I promoter examined by in vivo and in vitro footprinting.
Constitutive and metal-inducible protein:DNA interactions at the mouse metallothionein I promoter examined by in vivo and in vitro footprinting.
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通过体内和体外足迹检测小鼠金属硫蛋白 I 启动子的组成型和金属诱导型蛋白：DNA 相互作用。
DOI：
10.1101/gad.2.4.412
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发表时间：
1988
影响因子：
10.5
通讯作者：
Wold,B
中科院分区：
文献类型：
--
作者：
Mueller,PR;Salser,SJ;Wold,B
已经开发了一种高分辨率体内足迹法，并用于调查小鼠金属硫蛋白I（MT-I）启动子与基础水平转录和高水平金属诱导转录相关的蛋白质：DNA相互作用。该启动子及其相关的调控区在结构上是复杂的。它含有多个潜在的金属调节因子和其他转录因子，包括SP1和MLTF的结合位点。在几种情况下，潜在的识别位点重叠，这里报道的实验提供了一个视图的网站在体内使用。这些数据还显示了当细胞从基础水平表达转变为金属诱导表达时，蛋白质：DNA接触的模式如何变化。非诱导足迹模式由相互作用的基础元素，被认为是负责该基因的中度转录在没有添加金属。这些相互作用在金属诱导后保持不变。当MT-I表达增加暴露于锌或镉的细胞，观察到一个新的足迹模式。它包括基础相互作用和一组新的金属依赖足迹，这些足迹位于所有五个遗传定义的金属响应元件（MRE）MRE-A-MRE-E上。此外，这些数据确定了第六个可能的MRE，MRE-F，它显示出与其他MRE相似的硫酸二甲酯（DMS）足迹。
A method of high resolution in vivo footprinting has been developed and used to survey the mouse metallothionein I (MT-I) promoter for protein : DNA interactions associated with basal-level transcription and with high-level metal-induced transcription. This promoter and its associated regulatory region is structurally complex. It contains multiple potential binding sites for metal regulatory factors and for other transcription factors, including SP1 and MLTF. In several cases potential recognition sites overlap, and the experiments reported here provide a view of which sites are utilized in vivo. These data also show how the pattern of protein : DNA contacts changes when cells are shifted from basal-level expression to metal-induced expression. The noninduced footprint pattern consists of interactions at basal elements that are thought to be responsible for the moderate transcription of this gene in the absence of added metals. These interactions remain unchanged upon metal induction. When MT-I expression is increased by exposing cells to zinc or cadmium, a new footprint pattern is observed. It includes the basal interactions and a new set of metal-dependent footprints that are positioned over all five genetically defined metal responsive elements (MREs), MRE-A--MRE-E. In addition, these data identify a sixth probable MRE, MRE-F, which displays a dimethylsulfate (DMS) footprint similar to that at other MREs.
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