MECHANISMS OF YEAST TRANSCRIPTIONAL REGULATORS

这项工作的长期目标是了解，在精确的分子 术语，芽殖酵母中细胞类型特异性基因如何转录 并将从这些研究中发展出来的原则应用到更多的领域。 复杂的有机体该提案可分为三个基本问题： (l)同源异型结构域蛋白（以α 1和β 2为例）是如何 α 2）以高亲和力和特异性识别靶DNA序列？ (2)什么是一般转录的作用机制 阻遏系统（SSN 6/TUP 1）在酵母？(3)是 抑制机制和抑制机制保存在更多的 复杂的有机体？ 实验方法强调使用纯化的蛋白质， 生化实验来推断分子机制。遗传方法 在酵母中被利用（I）来测试从生物化学开发的模型， 研究和（2）识别阻遏系统的缺失成分。 最后，酵母中的功能互补和基于PCR的策略 用于鉴定和分析阻遏系统的组分 在其他生物体中。 鉴于基因调控蛋白的高度保守性， 在所有真核生物中， 本书所述的酵母蛋白质的许多原理， 建议将适用于其他设置。基本了解 细胞特化背后的分子事件不仅提供了 理解流程如何失败的框架，但也提供了 设计治疗策略的基础和知识， 干预

The long term goal of this work is to understand, in precise molecular terms, how cell-type specific genes in budding yeast are transcriptionally repressed and to apply the principles developed from these studies to more complex organisms. The proposal can be divided into three basic questions: (l) How do pairs of homeodomain proteins (exemplified by alpha1 and alpha2) recognize with high affinity and specificity target DNA sequences? (2) What is the mechanism of action of a general transcriptional repression system (SSN6/TUP1) in yeast? (3) Are the components of the repression machinery and the mechanisms of repression conserved in more complex organisms? The experimental approaches emphasize the use of purified proteins and biochemical experiments to deduce molecular mechanisms. Genetic approaches in yeast are utilized (I) to test models developed from the biochemical studies and (2) to identify missing components of the repression system. Finally, complementation for function in yeast and PCR-based strategies are utilized to identify and analyze components of the repression systems in other organisms. Given the high degree of conservation of gene regulatory proteins and general transcription factors among all 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.