Molecular Analysis of a Yeast Transcriptional Regulator

简介(由申请人提供)：建议的长远目标 本项目旨在阐明Mot1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 抄写。Mot1是进化保守的一个大家族的成员 核ATPase(Snf2/Swi2家族)参与转录、DNA修复、 和重组。人类Snf2/Swi2相关蛋白复合体的缺陷是 已知会导致某些儿科癌症，Cockayne综合征， A型地中海贫血，是最常见的X连锁智力低下。尽管 Snf2/Swi2家族成员的普遍存在及其分子机制 这些蛋白质的作用不是很详细，也不是很清楚 这些蛋白质中的许多在体内扮演着什么角色。 Mot1的S ATPase活性是激活或抑制转录所必需的 体内的特定基因。与其作为抑制者的角色一致，Mot1可以 三磷酸腺苷依赖蛋白中TBP-DNA复合体的解离 反应。Motl依赖于ATP的转录激活机制是 未知。将使用生化、分子生物学和遗传学方法 为了定义Mot1如何在体内调节转录，以及Mot1如何响应 确定了特定的推动者。这些方法还将用于 了解Mot1对ATP的水解如何导致TBP/DNA中断。建议数 对Mot1功能的分析将有助于更好地理解Mot1的作用 Motl在转录调控中的作用以及更好地理解 Snf2/Swi2家族成员的一般功能。

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.