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Study of molecular structural and dynamical aspects of transcriptional regulatory factors

转录调控因子的分子结构和动力学方面的研究

基本信息

批准号：
16370051
负责人：
OAGATA Kazuhiro
金额：
$ 9.79万
依托单位：
Yokohama City University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2005
项目状态：
已结题

项目摘要

The goal of this project is to elucidate molecular mechanisms for regulating the enhanceosome formation, particularly, stabilization of transcription factors bound to an enhancer region of a gene.We investigated some complexes containing a transcriptional regulatory factor, Runx1, which plays a pivotal role in hematopoiesis and is involved in acute leukemias. The DNA binding activity of Runxl is enhanced allosterically by non-DNA binding heterodimeric partner, CBFβ. Previously, we showed that there was no significant structural change of Runxl upon binding of CBFβ when a crystal structure of Runx1-DNA complex was compared with that of Runx1-CBFβ-DNA complex. It promoted us to focus on molecular fluctuations of Runx1. Magnetic relaxation measurements using NMR demonstrated that some regions of Runx1 are significantly fluctuated without CBFβ, while the fluctuations of Runx1 became reduced upon binding of CBFβ. We proposed that regulations of molecular functions are accomplished by alteri … More ng molecular fluctuations.Runxl is also involved in transcriptional regulation of the T cell antigen receptor a chain (TCRα) together with another transcriptional regulatory factor, Etsl. The DNA binding activity of Etsl is controlled by an N-terminal flanking region of ETS domain, referred exon VII. The exon VII inhibits the DNA binding activity of Ets1 while the inhibition is released by an adjacently placed Runxl. Etsl is not capable of binding the enhancer upon phosphorylation of exon VII even if Runxl binds to the enhancer. We investigated the molecular details of the complex composed of Runxl and Etsl in the TCRα enhancer using X-ray crystallography, EMSA and the luciferase reporter gene assay. We identified which regions of Etsl are responsible for cooperative DNA binding with Runxl and the impact of the exon VII phosphorylation. Further structural analyses might be required to unveil overall aspect of the regulatory systems. We will also study about fluctuations of Runxl upon Etsl binding. Less

该项目的目标是阐明调节增强体形成的分子机制，特别是与基因增强子区域结合的转录因子的稳定性。我们研究了一些含有转录调节因子Runx1的复合物，Runx1在造血中起关键作用，并参与急性白血病。Runxl的DNA结合活性被非DNA结合的异二聚体伙伴CBFβ变构增强。在此之前，我们发现Runx1-DNA复合物的晶体结构与runx1 - cbf - β- dna复合物的晶体结构相比，Runx1-DNA复合物与cbf - β结合后，Runxl的结构没有明显变化。这促使我们关注Runx1的分子波动。利用核磁共振磁弛豫测量表明，Runx1的一些区域在没有CBFβ的情况下有明显的波动，而与CBFβ结合后Runx1的波动减小。我们提出分子功能的调节是通过分子的波动来完成的。Runxl还与另一个转录调控因子Etsl一起参与T细胞抗原受体a链（TCRα）的转录调控。Etsl的DNA结合活性由ETS结构域n端侧翼区域控制，称为外显子VII。外显子VII抑制Ets1的DNA结合活性，而邻近的Runxl则释放这种抑制。即使Runxl与增强子结合，Etsl也不能在外显子VII磷酸化时与增强子结合。我们利用x射线晶体学、EMSA和荧光素酶报告基因实验研究了TCRα增强子中Runxl和Etsl复合物的分子细节。我们确定了Etsl的哪些区域负责与Runxl的合作DNA结合以及外显子VII磷酸化的影响。可能需要进一步的结构分析，以揭示监管体系的整体方面。我们还将研究Runxl在Etsl结合后的波动。少

项目成果

期刊论文数量（10）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Cyclin-dependent kinase (CDK) phosphorylation destabilizes somatic Weel via multiple pathways.

细胞周期蛋白依赖性激酶 (CDK) 磷酸化通过多种途径破坏体细胞 Weel 的稳定性。

DOI：
发表时间：
2005
期刊：
Proc.Natl.Acad.Sci.USA 102
影响因子：
0
作者：
Nishino;N.;Shivashimpi;G.M.;Soni;P.B.;Bhuiyan;M.P.I.;Kato;T.;Maeda;S.;Nishino;T.G.;Yoshida;M.;N.Watanabe et al.
通讯作者：
N.Watanabe et al.

Assembly of transcriptional regulatory factors, c-Myb and C/EBP β, from separated sites on a promoter

从启动子上的不同位点组装转录调节因子 c-Myb 和 C/EBP β