在TFIIH中PH域的结构多态性
Masahiko Okuda1, Yoshifumi Nishimura1,2
1Graduate School of Medical Life Science, Yokohama City University, 1-7-29 Suehiro-cho, Tsurumi-ku, Yokohama 230-0045, Japan.
通用转录因子TFIIH的斑纹蛋白同质 (PH) 域与人类PH域具有结构上的相似性,而不是芽酵母PH域. 这种PH域在不同的酵母和甲基动物物种中以不同的方式与标蛋白相互作用.
科学领域:
- 分子生物学分子生物学
- 结构生物学 结构生物学
- 生物化学 生物化学
背景情况:
- 一般转录因子TFIIH对于转录,DNA修复和细胞循环调节在真核生物中至关重要.
- 在TFIIH子单元 (如人体p62和酵母Tfb1) 中的斑纹蛋白同质 (PH) 域调解了特定DNA位点的招募.
- 的PH域结构上是不同的,以前只有Saccharomyces cerevisiae的PH域结构是已知的.
研究的目的:
- 使用核磁共振 (NMR) 来确定Schizosaccharomyces pombe (裂变酵母) Tfb1 (spPH) 的PH域的结构.
- 将spPH的结构和目标结合特性与已知的人类 (hPH) 和芽酵母 (scPH) 领域进行比较.
- 为了阐明spPH与其相互作用伙伴spTfa1和spRhp41的结合模式.
主要方法:
- 核磁共振 (NMR) 光谱法用于确定spPH域的3D结构.
- 生物信息学分析被用来比较氨基酸序列的同一性,并预测目标结合部位.
- 进行了化学转移干扰实验,以确定spPH及其标蛋白之间的结合接口.
主要成果:
- spPH域表现出与人类PH域 (hPH) 相比与芽酵母PH域 (scPH) 更密切相关的架构,尽管与scPH的序列相同性更高.
- 特定结合的关键残留物,在hPH中确定,在spPH中存在,表明功能重要性得到保留.
- spPH与spTfa1和spRhp41通过不同的结合方式在类似但独特的表面上相互作用,与hPH和scPH相互作用不同.
结论:
- TFIIH的PH域显示结构和功能可塑性,裂变酵母的PH域与甲状动物对应物相似,而不是与芽酵母相似.
- 多态结合模式突显了TFIIH PH域在与不同真核细胞系的多种标蛋白相互作用时的适应性.
- 这项研究揭示了对转录和DNA修复中TFIIH PH域的进化分歧和保存的功能元素的新见解.
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