Mechanisms of Yeast Transcriptional Regulators

超出提供的空间。这项提议有两个长期目标。首先是用精确的分子术语理解高度保守的基因转录抑制物的作用机制。这项工作将与酿酒酵母Sn6-Tupl抑制子一起进行，并强调使用纯化的蛋白质进行生化和结构方法，旨在推断分子机制。酿酒酵母中的遗传方法，包括广泛使用微阵列技术，将被用于测试来自生化实验的模型，并作为探索工具。第二个长期目标是描述酿酒酵母和几种相关酵母中整个SSN6-Tup1调控网络，包括所有利用SSN6-Tup1的DNA结合蛋白和由每个DNA结合蛋白控制的全套基因。这项工作的最终目标是理解由高度保守的组件组成的调节电路如何在进化时间尺度上变得专门化。鉴于基因调控蛋白和一般转录因子在真核生物中的高度保守性，似乎本提案中为酵母蛋白开发的许多原理将适用于其他环境。对细胞特化背后的分子事件的基本了解不仅为理解该过程如何失败提供了一个框架，而且还为基于干预的治疗策略的设计提供了基础和知识。表演网站========================================Section End===========================================

EXCEED THE SPACE PROVIDED. This proposal has two long-term goals. The first is an understanding, in precise molecular terms, of the mechanism of action of highly conserved represser of gene transcription. This work will be carried out with the S. cerevisiae Ssn6-Tupl represser and emphasizes the use of purified proteins for biochemical and structural approaches designed to deduce molecular mechanisms. Genetic approaches in S. cerevisiae, which include the extensive use of microarray technologies will be used to test models derived from biochemical experiments and as exploratory tools. The second long-term goal is a description of the entire Ssn6-Tupl regulatory network in S. cerevisiae and several related yeasts, including all the DNA-binding proteins that utilize Ssn6-Tupl and the complete sets of genes controlled by each DNA-binding protein. The ultimate goal of this work is an understanding of how regulatory circuits that are composed of highly conserved components become specialized over evolutionary time scales. Given the high degree of conservation of gene regulatory proteins and general transcription factors among eucaryotes, it seems likely that many of the principles developed for the yeast proteins covered in this proposal will apply in other settings. A basic understanding of the molecular events underlying cell specialization provides not only a framework for understanding how the process can fail, but also provides the substrates and knowledge to design therapeutic strategies based on intervention. PERFORMANCE SITE ========================================Section End===========================================