酵母TATA盒结合因子TFIID的结构功能关系的分析
M Horikoshi1, T Yamamoto, Y Ohkuma
1Laboratory of Biochemistry and Molecular Biology, Rockefeller University, New York, New York 10021.
Cell
|June 29, 1990
概括
研究人员在Saccharomyces cerevisiae TFIID中发现了一个大型C端区域 (残留物63-240),该区域负责TATA盒结合和转录. 这一区域在各个物种中都存在,TFIID将DNA作为单体结合在一起.
科学领域:
- 分子生物学分子生物学
- 生物化学 生物化学
- 遗传学 是一个遗传学.
背景情况:
- 转录因子II D (TFIID) 是真核生物中关键的一般转录因子.
- 了解TFIID域的结构功能关系对于阐明转录调节至关重要.
- 之前的研究表明,TFIID的C端区域内保留了结构图案.
研究的目的:
- 系统地绘制Saccharomyces cerevisiae TFIID的功能域,负责DNA结合和转录活动.
- 调查保存的C终端区域在TFIID功能中的作用.
- 在DNA结合过程中确定TFIID的寡合状态.
主要方法:
- 构建和体外表达S. cerevisiae TFIID的N端,C端和内部删除突变体.
- 用DNA移动性转移测试来评估TATA盒结合活性.
- 试管体内转录测试用于评估基底水平的转录活性.
- 删除突变体的共转化研究,以确定结合性静脉测量.
主要成果:
- 鉴定出一个重要的C端区域 (63-240残留物) 含有负责TATA盒结合和基底转录的域.
- 这一功能关键区域包含了先前注意到的结构动图,并且在物种之间高度保存.
- 删除突变共翻译研究表明,TFIID以单体形式与DNA结合.
结论:
- S. cerevisiae TFIID的大C终端区域 (63-240) 对于其DNA结合和转录激活功能至关重要.
- TFIID与DNA的单体结合对其组装和在预启动复合体中的作用有影响.
- 这些发现有助于更深入地了解TFIID结构,保存及其在真核细胞转录启动中的作用机制.
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