GAL4衍生物单独和与哺乳动物激活剂在体外协同作用
Y S Lin1, M F Carey, M Ptashne
1Department of Biochemistry and Molecular Biology, Harvard University, Cambridge, Massachusetts 02138.
Cell
|August 26, 1988
概括
酵母GAL4蛋白衍生物在体外激活哺乳动物基因转录. 有效的激活需要特定的结合部位放置或与哺乳动物激活剂的协同作用,揭示了新的体外特定激活领域.
科学领域:
- 分子生物学分子生物学
- 基因规则 基因规则
- 蛋白质工程是指蛋白质工程.
背景情况:
- 酵母转录激活剂GAL4是一种经过充分研究的转录因子.
- 了解真核生物基因转录机制对于各种生物过程至关重要.
研究的目的:
- 在哺乳动物系统中研究GAL4衍生物的体外转录激活能力.
- 确定GAL4结合部位位置和与其他激活剂的相互作用对转录刺激的影响.
主要方法:
- 来自大肠杆菌的GAL4衍生物的合成和净化.
- 在体外转录试验中,使用HeLa细胞核提取物和修饰的腺病毒E4基因模板进行了转录试验.
- 与E4 TATA盒相比,GAL4结合点位置的系统变化.
主要成果:
- GAL4衍生物有效地刺激了HeLa核提取物中的腺病毒E4基因转录.
- 当GAL4结合部位位于TATA盒的上游时,就会发生最佳刺激.
- 遥远的GAL4位点需要近邻哺乳动物激活器位点进行协同刺激,突出显示了依赖上下文的激活.
结论:
- GAL4衍生物可以在异种哺乳动物系统中作为转录激活剂发挥作用.
- 该研究确定了GAL4中的第三个激活域,仅在体外功能.
- 结果提供了对转录激活机制的模块化和上下文依赖性质的见解.
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