Chemical Tools for Regulating Eukaryotic Transcription

描述：（由申请人提供）人类基因组的测序偶联 随着蛋白质组学和基因组学的不断进步， 了解基因内容与疾病之间的关系。作为 因此，基于转录的治疗方法的发展， 选择性重编程病变细胞中的异常基因表达是一种 越来越有吸引力的目标。然而，仍然存在一些基本问题， 围绕转录的调节机制以及 对这种非天然转录的特异性和活性的要求 因素这些问题中的许多已被证明是难以解决的遗传或 生物化学手段。本文描述了一种用于生成 独特的化学工具，将用于解决的基本问题， 转录调控这些工具将进一步作为分子靶点 长期发展基于转录的疗法。 酿酒酵母或芽殖酵母将作为我们的模型系统， 与哺乳动物系统的机制同源性，基因组的可用性， 信息，并且因为酵母特异性转录激活因子和/或 阻遏物将作为抗真菌剂的优良候选物。 为了实现这些目标，将选择转录调节子 从筛选与介体复合物结合的肽库中 组分Gall 1.一系列生化技术将被用来 表征如此选择的所有配体的结合亲和力和特异性。 体外转录实验将用于功能上 将配体表征为转录的激活剂或抑制剂， 用于探测转录调控的机制问题 在设计用于绘制重要功能蛋白质-蛋白质图谱的实验中， 相互作用，并探测转录激活中协同作用的起源。 由于酿酒酵母是后生动物的一个很好的模型系统， 由此获得的数据将大大有助于不断发展的模型， 真核转录激活。此外，化学调节剂 在整个项目中开发的将作为实现 长期目标的设计和调查基于转录 治疗学正在研究的蛋白质Galil是一种酵母特异性蛋白质， 转录装置的组成部分;因此， 通过与Gall 1的特异性相互作用发挥其功能， 在酵母中，为抗真菌剂的开发提供了一种机制。

DESCRIPTION: (provided by applicant) The sequencing of the human genome coupled with continued advances in proteomics and genomics have led to an unprecedented understanding of the relationship between genetic content and disease. As a consequence, the development of transcription-based therapeutics that would selectively reprogram aberrant gene expression in diseased cells is an increasingly attractive goal. There remain, however, fundamental questions surrounding the regulatory mechanism of transcription as well as the requirements for specificity and activity of such non-natural transcription factors. Many of these questions have proven intractable by genetic or biochemical means alone. Described herein is an approach for the generation of unique chemical tools that will be used for addressing basic questions of transcriptional regulation. These tools will further serve as molecular targets for the long-term development of transcription-based therapeutics. Saccharomyces cerevisiae or budding yeast will serve as our model system due to the mechanistic homology with mammalian systems, the availability of genomic information, and because yeast-specific transcriptional activators and/or repressors will serve as excellent candidates for anti-fungal agents. In order to accomplish these goals, transcriptional regulators will be selected from libraries of peptides screened for binding to the Mediator complex component Gall 1. A battery of biochemical techniques will be used to characterize the binding affinity and specificity of all ligands thus selected. In vitro transcription experiments will be employed to functionally characterize the ligands as activators or inhibitors of transcription that will be used to probe mechanistic questions surrounding transcriptional regulation in experiments designed to map functionally important protein-protein interactions and to probe the origin of synergy in transcriptional activation. As Saccharomyces cerevisiae is an excellent model system for metazoans, the data thus obtained will significantly contribute to the evolving model of eukaryotic transcriptional activation. Furthermore, the chemical regulators developed throughout the project will serve as the basis for achievement of the long-term goal of the design and investigation of transcription-based therapeutics. The protein under investigation, Galil, is a yeast-specific component of the transcriptional apparatus; thus artificial regulators that exert their function via specific interactions with Gall 1 will function only in yeast, providing a mechanism for the development of anti-fungal agents.