Chemical Tools for Regulating Eukaryotic Transcription

调节真核转录的化学工具

• 批准号: 6723782
• 负责人: ANNA K. MAPP
• 金额: $ 21.14万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-04-01 至 2007-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6723782
• 关键词: Saccharomyces cerevisiae; biotechnology; functional /structural genomics; fungal genetics; fungal proteins; genetic transcription; high throughput technology; peptide library; protein binding; protein protein interaction; transcription factor

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.

描述：（由申请人提供）人类基因组的测序耦合 随着蛋白质组学和基因组学的持续进展，导致了前所未有的 了解遗传含量与疾病之间的关系。作为 结果，开发基于转录的治疗剂 在患病细胞中选择性重编程异常基因表达是一种 越来越有吸引力的目标。但是，仍然存在基本问题 围绕转录的调节机制以及 这种非天然转录的特异性和活动的要求 因素。这些问题中的许多已证明是通过遗传或 生化意味着单独的意思。此处描述的是一种生成的方法 独特的化学工具将用于解决 转录调节。这些工具将进一步用作分子目标 用于基于转录的治疗剂的长期发展。 酿酒酵母或萌芽的酵母将作为我们的模型系统 与哺乳动物系统的机械同源性，基因组的可用性 信息，以及因为酵母特异性的转录激活剂和/或 阻遏物将作为抗真菌药物的出色候选者。 为了实现这些目标，将选择转录调节器 从筛选的肽库中，与中介体复合物结合 组件胆汁1。将使用一系列生化技术 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 将配体的特征为激活剂或转录抑制剂 用于探测围绕转录调节的机理问题 在旨在绘制功能上重要蛋白质蛋白的实验中 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出色模型系统，因此 因此获得的数据将显着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是实现的基础 基于转录的设计和研究的长期目标 疗法。正在研究的蛋白质Galil是酵母特异性的 转录设备的组成部分；因此，人工调节器 通过与GALL 1的特定相互作用发挥其功能仅将起作用 在酵母中，提供了一种抗真菌剂的机制。