Chemical Tools for Regulating Eukaryotic Transcription

描述：(申请人提供)结合的人类基因组测序 随着蛋白质组学和基因组学的不断进步，导致了前所未有的 了解基因含量和疾病之间的关系。作为一名 结果，基于转录的疗法的发展将 选择性地重新编程疾病细胞中异常基因的表达是一种 越来越有吸引力的目标。然而，仍然存在一些根本性的问题 围绕转录的调节机制以及 对这种非自然转录的特异性和活性的要求 各种因素。这些问题中的许多都被证明是由于遗传或 生化手段仅指。这里描述的是一种用于生成 独特的化学工具将用于解决以下基本问题 转录调控。这些工具将进一步成为分子靶标。 用于基于转录的疗法的长期发展。 酿酒酵母或萌芽酵母将作为我们的模型系统，因为 与哺乳动物系统的机械同源性，基因组的可用性 信息，并且因为酵母特定的转录激活子和/或 抑制物将成为抗真菌药物的极佳候选者。 为了实现这些目标，将选择转录调控因子 从筛选的与介体复合体结合的多肽文库中 一组生化技术将被用来 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 将配体表征为转录的激活剂或抑制物 用来探索围绕转录调控的机械性问题 在设计用于绘制具有重要功能的蛋白质的实验中 相互作用，并探讨转录激活中协同作用的起源。 由于酿酒酵母是后生动物的优秀模式系统， 这样获得的数据将极大地促进 真核转录激活。此外，化学监管机构 整个项目的开发将作为实现以下目标的基础 基于转录的设计和研究的长期目标 治疗学。研究中的蛋白质Galil是一种酵母特有的蛋白质 转录装置的组成部分；因此，人工调节器 通过与Gall 1的特定交互来发挥其功能仅起作用 在酵母中，为抗真菌药物的发展提供了一种机制。

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.