MOLECULAR ANALYSIS OF A YEAST TRANSCRIPTIONAL REGULATOR

酵母转录调节因子的分子分析

• 批准号: 2024315
• 负责人: David T. Auble
• 金额: $ 20.66万
• 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 2002-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2024315
• 关键词: DNA binding protein; DNA footprinting; adenosinetriphosphatase; antibody; crosslink; enzyme activity; fungal genetics; gel mobility shift assay; genetic library; genetic promoter element; intermolecular interaction; laboratory rabbit; nucleic acid sequence; protein purification; protein structure function; site directed mutagenesis; transcription factor; western blottings; yeasts

The long-term objectives of the proposed project are to elucidate the mechanism of action and in vivo function of MOT1, an essential yeast protein that regulates transcription by dissociating TATA-binding protein (TBP)-DNA complexes in an ATP-dependent manner. MOT1 is a member of a large and growing family of evolutionarily conserved nuclear proteins (the SNF2/SWI2 family) involved in transcription, DNA repair, recombination, and chromosome segregation. Mutation of SNF2/SWI2 family members in humans have been identified as the causes of Cockayne's syndrome, X-linked mental retardation, and alpha-thalassemia. 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 most of these proteins play in vivo. Biochemical, molecular biological and genetic approaches will be used to define in molecular detail how MOT1 interacts with TBP, and how its ATPase is utilized to drive TBP-DNA complex dissociation. These approaches will also be used to identify other proteins that MOT1 interacts with in vivo and to define the parameters that determine the magnitude of MOT1's effect on the expression of individual genes in vivo. The proposed analysis of MOT1 function will lead to a better understanding of the role of MOT1 as well as a better understanding of the functions of SNF2/SWI2 family members in general. The proposed work will also provide a framework for the analysis of other SNF2/SWI2 family members whose activities are currently poorly understood.

拟议项目的长远目标是阐明 一种必需酵母MOT 1的作用机制和体内功能 通过分离TATA结合蛋白来调节转录的蛋白质 （TBP）-DNA复合物以ATP依赖的方式。 MOT 1是一个 进化上保守的核蛋白的一个大的和不断增长的家族（ SNF 2/SWI 2家族）参与转录、DNA修复、重组， 和染色体分离。 SNF 2/SWI 2家族成员的突变 人类已被确定为Cockayne综合征的原因，X连锁 智力迟钝和α-地中海贫血 尽管无处不在的 SNF 2/SWI 2家族成员的发生、作用的分子机制 这些蛋白质的细节并不清楚，也不知道它们是什么。 这些蛋白质中的大多数在体内发挥作用。 生物化学、分子生物学和遗传学方法将用于 在分子上详细定义MOT 1如何与TBP相互作用，以及它的ATP酶如何与TBP相互作用。 用于驱动TBP-DNA复合物解离。 这些办法将 也可用于鉴定MOT 1在体内与之相互作用的其他蛋白质 并定义确定MOT 1效应大小的参数 对个体基因在体内表达的影响。 拟议的分析 MOT 1的功能将有助于更好地理解MOT 1的作用， 以及对SNF 2/SWI 2家族功能的进一步了解 成员一般。 拟议的工作还将提供一个框架， 对其他SNF 2/SWI 2家族成员的分析， 目前了解甚少。