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Mechanisms of chromatin organization in genome dictated by DNA structural properties.

基因组中染色质组织机制由 DNA 结构特性决定。

基本信息

批准号：
14572079
负责人：
SHIMIZU Mitsuhiro
金额：
$ 2.24万
依托单位：
Meisei University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2003
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14572079/
关键词：
chromatin nucleosome DNA structure transcriptional regulation repetitive sequences budding yeast genome

项目摘要

Gene expression in eukaryotic cells involves the interaction of proteins with DNA packaged into nucleosomes. Nucleosomes are often located at distinct positions in promoter regions to regulate transcription. Thus, the factors that influence the nucleosome positioning are essential to elucidating the mechanism of gene expression. In this project, we have examined the role of DNA structures in chromatin organization and its effect on gene expression in vivo.1.Effect of DNA structures on nucleosome positioning in vivo.We have examined effect of simple repeating sequences that can form alternative DNA conformations on nucleosome positioning using yeast minichromosome system. The repeating sequences are categorized into 4 groups in terms of nucleosome-promoting and nucleosome-disrupting properties.2.Effect of chromatin alteration through DNA structures on repression in yeast genome.In Saccharomyces cerevisiae, nucleosomes are precisely positioned in the promoters of a cell-specific genes such as STE6 and BAR1. Here, we have demonstrated that chromatin alteration by intrinsic DNA structures disrupts repression of BAR1 in genome and of STE6-lacZ reporter plasmids in a cells. Therefore, proper nucleosome positioning is essential for regulation of cell type-specific transcription and cell type identity.3.Effect of triplet repeat sequences on chromatin organization.We have shown that triplet repeat sequences that cause human hereditary diseases affect nucleosome organization in yeast minichromosomes. Their effect on gene expression is associated with their features on chromatin structure. The sticky DNA formation at longer GAA repeats from Friedreich's ataxia was also investigated in yeast minichromosomes.In summary, intrinsic DNA structures play important roles in nucleosome organization in vivo. Our studies also emphasizes that chromatin alteration through DNA structures would make it possible to modulate gene expression artificially in eukaryotes.

真核细胞中的基因表达涉及蛋白质与包装到核小体中的 DNA 的相互作用。核小体通常位于启动子区域的不同位置来调节转录。因此，影响核小体定位的因素对于阐明基因表达机制至关重要。在这个项目中，我们研究了DNA结构在染色质组织中的作用及其对体内基因表达的影响。1.DNA结构对体内核小体定位的影响。我们利用酵母微染色体系统研究了可以形成替代DNA构象的简单重复序列对核小体定位的影响。根据核小体促进和核小体破坏特性，重复序列分为 4 组。2.通过 DNA 结构改变染色质对酵母基因组抑制的影响。在酿酒酵母中，核小体精确定位在细胞特异性基因（如 STE6 和 BAR1）的启动子中。在这里，我们证明了内在 DNA 结构引起的染色质改变会破坏基因组中 BAR1 和细胞中 STE6-lacZ 报告质粒的抑制。因此，正确的核小体定位对于调节细胞类型特异性转录和细胞类型身份至关重要。3.三联体重复序列对染色质组织的影响。我们已经证明，引起人类遗传性疾病的三联体重复序列影响酵母微染色体中的核小体组织。它们对基因表达的影响与其对染色质结构的特征有关。弗里德赖希共济失调导致的较长 GAA 重复序列中粘性 DNA 的形成也在酵母微型染色体中进行了研究。总之，内在 DNA 结构在体内核小体组织中发挥着重要作用。我们的研究还强调，通过 DNA 结构改变染色质将使在真核生物中人为调节基因表达成为可能。

项目成果

期刊论文数量（46）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Yeast Ume6p repressor permits activator binding but restricts TATA-box binding protein association with the HOP1 promoter.

酵母 Ume6p 阻遏蛋白允许激活剂结合，但限制 TATA-box 结合蛋白与 HOP1 启动子的关联。

DOI：
发表时间：
2003
期刊：
Nucleic Acids Res. 31
影响因子：
0
作者：
Mitsuhiro Shimizu;Keiko Takahashi;Teresa Lamb;Heisaburo Shindo;Aaron P.Mitchell
通讯作者：
Aaron P.Mitchell

Separation and recovery of DNA fragments by electrophoresisthrough a thermoreversible hydrogel of multipliedblock copolymers composed of poly(ethylene oxide) and poly(propylene oxide).

通过由聚环氧乙烷和聚环氧丙烷组成的倍增嵌段共聚物的热可逆水凝胶进行电泳分离和回收 DNA 片段。