Molecular Analysis of a Yeast Transcriptional Regulator

酵母转录调节因子的分子分析

• 批准号: 6625613
• 负责人: David T. Auble
• 金额: $ 28.7万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-05-01 至 2006-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6625613
• 关键词: DNA binding protein; DNA footprinting; adenosinetriphosphatase; enzyme activity; fluorescence polarization; fungal genetics; fungal proteins; gel mobility shift assay; gene induction /repression; gene mutation; genetic library; genetic promoter element; immunoprecipitation; intermolecular interaction; laboratory rabbit; nucleic acid sequence; protein purification; protein structure function; site directed mutagenesis; transcription factor; western blottings; yeasts

DESCRIPTION (provided by applicant): The long-term objectives of the proposed project are to elucidate the mechanism of action and in vivo function of Mot1, an essential yeast transcriptional regulator that can activate or repress transcription. Mot1 is a member of a large family of evolutionarily conserved nuclear ATPases (the Snf2/Swi2 family) involved in transcription, DNA repair, and recombination. Defects in human Snf2/Swi2-related protein complexes are known to contribute to certain pediatric cancers, Cockayne's Syndrome, a-thalassemia, and the most common form of X-linked mental retardation. Despite the ubiquitous occurrence of Snf2/Swi2 family members, the molecular mechanisms of action of these proteins are not understood in detail, nor is it understood what roles many of these proteins play in vivo. Mot1 's ATPase activity is required to activate or repress transcription of specific genes in vivo. Consistent with its role as a repressor, Mot1 can dissociate TATA-binding protein (TBP)-DNA complexes in an ATP-dependent reaction. Motl's mechanism of ATP-dependent transcriptional activation is unknown. Biochemical, molecular biological, and genetic approaches will be used to define how Mot1 regulates transcription in vivo and how the Mot1 response of particular promoters is determined. These approaches will also be used to understand how ATP hydrolysis by Mot1 drives TBP/DNA disruption. The proposed analysis of Mot1 function will lead to a better understanding of the role of Motl in transcriptional control as well as a better understanding of the functions of Snf2/Swi2 family members in general.

描述（由申请人提供）：拟议的长期目标 项目是阐明Mot 1的作用机制和体内功能， 一种重要的酵母转录调节因子，可以激活或抑制 转录。Mot 1是一个进化上保守的大家族成员， 核ATP酶（Snf 2/Swi 2家族）参与转录，DNA修复， 和重组。人Snf 2/Swi 2相关蛋白复合物的缺陷是 已知会导致某些儿科癌症，科凯恩综合征， α-地中海贫血和最常见的X连锁智力低下。尽管 Snf 2/Swi 2家族成员的普遍存在， 这些蛋白质的作用机制尚未被详细了解， 这些蛋白质在体内起着什么样的作用 Mot 1的ATP酶活性是激活或抑制转录所必需的。 体内的特定基因与其作为阻遏物的作用一致，Mot 1可以 在ATP依赖性的条件下解离TATA结合蛋白（TBP）-DNA复合物 反应Motl的ATP依赖性转录激活机制是 未知将采用生物化学、分子生物学和遗传学方法 确定Mot 1如何在体内调节转录，以及Mot 1如何响应 确定特定的启动子。这些方法还将用于 了解Mot 1的ATP水解如何驱动TBP/DNA破坏。拟议 对Mot 1功能的分析将有助于更好地理解 Motl在转录调控中的作用以及对Motl在转录调控中的作用的更好理解。 Snf 2/Swi 2家族成员的功能。