Characterization of a myeloid-specific enhancer of the chicken lysozyme gene. Major role for an Ets transcription factor-binding site.
Characterization of a myeloid-specific enhancer of the chicken lysozyme gene. Major role for an Ets transcription factor-binding site.
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鸡溶菌酶基因的骨髓特异性增强子的表征。
DOI：
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发表时间：
1994
影响因子：
4.8
通讯作者：
W. Strätling
中科院分区：
文献类型：
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作者：
B. Ahne;W. Strätling
溶菌酶基因在鸡巨噬细胞中的表达受位于转录起始位点上游2.7个酶的增强子调控。该增强子的组织通过体外测定（DNA酶I足迹法、硫酸二甲酯甲基化保护和带移测定）和体内足迹法实验进行分析。结果表明，增强子包含四个区域（I-IV）的蛋白质-DNA相互作用。首先，瞬时基因转移实验表明，区域II对增强子功能贡献最大。该区域含有巨噬细胞和B细胞特异性转录因子PU. 1的识别位点，PU. 1是Ets转录因子家族的成员。该位点在增强子中起主要作用，因为该位点的突变消除了增强子活性。第二，在竞争带移位实验中，我们表明，只有骨髓特异性复合物在区域I和II与核因子形成的PU。1识别位点的Ets转录因子家族的成员。第三，在体内足迹实验中，蛋白质-DNA相互作用的唯一标志是与PU. 1识别位点3 '相邻的硫酸二甲酯-高反应性鸟嘌呤。
Expression of the lysozyme gene in chicken macrophages is regulated by an enhancer located 2.7 kilobases upstream of the transcription start site. The organization of this enhancer was analyzed by in vitro assays (DNase I footprinting, dimethyl sulfate methylation protection, and band shift assays) and in vivo footprinting experiments. The results show that the enhancer contains four regions (I-IV) of protein-DNA interactions. First, transient gene transfer experiments demonstrate that region II contributes most to the enhancer function. This region contains a recognition site for the macrophage- and B cell-specific transcription factor PU.1, a member of the Ets transcription factor family. This site plays a major role in the enhancer since a mutation of this site abolishes enhancer activity. Second, in competition band shift experiments, we show that the only myeloid-specific complexes in regions I and II with nuclear factors are formed on the PU.1 recognition site by a member of the Ets transcription factor family. Third in in vivo footprinting experiments, the only sign of a protein-DNA interaction is a dimethyl sulfate-hyperreactive guanine 3'-adjacent to the PU.1 recognition site.