Eukaryotic Gene Regulation as Studied in Yeast

在酵母中研究的真核基因调控

• 批准号: 7195022
• 负责人: MARK S PTASHNE
• 金额: $ 71.9万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 1983
• 资助国家: 美国
• 起止时间: 1983-07-01 至 2010-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7195022
• 关键词: Accounting; Adhesives; Affect; Appearance; Base Pairing; Binding; Binding Sites; Biological Assay; Biological Process; Books; Cells; Chromatin Loop; Complex; DNA; DNA Binding; DNA-Binding Proteins; Dependency; Development; Drosophila bicoid protein; Eukaryota; Eukaryotic Cell; Excision; Funding; Galactose; Gene Activation; Gene Expression; Gene Expression Regulation; Genes; Genetic Transcription; Grant; Histones; Light; Location; Mammalian Cell; Measures; Mediating; Mediator of activation protein; Memory; Methods; Modeling; Nucleosomes; Organism; Paper; Peptides; Phase; Positioning Attribute; Process; Property; Proteins; Proteolysis; Rate; Recruitment Activity; Resolution; Series; Signal Transduction; Site; Surface; Testing; Time; Touch sensation; Transcription Coactivator; Transcriptional Activation; Ursidae Family; Work; Yeasts; gene induction; mutant; promoter; research study; response; zygote

DESCRIPTION (provided by applicant): Regulation of gene expression underlies disparate biological processes, including development of an organism from a fertilized egg, the response of organisms to extra-cellular signals, and the ingraining of memories. The grant requests funds to continue our analyses of mechanisms of gene regulation that will help us understand the normal processes and shed light on how they can go awry. The proposed studies use primarily yeast cells, a eukaryote in which many aspects of gene regulation are closely related to gene regulation in mammalian cells. In a series of papers and books we have developed a model - called the 'recruitment' model - for how transcriptional regulators determine whether any given gene will be activated or repressed. We propose here a series of experiments that probe and test this model in various ways. We have developed methods to measure the appearance, at a specific gene, of various protein complexes as they are recruited by a DNA-bound activator. Over 100 proteins must be brought to the gene for transcription to proceed, and we are using our methods to study how this large complex is formed stepwise. In a related development, we now can assay with great accuracy the disposition of nucleosomes on DNA, and we are studying how nucleosome positioning and removal is effected by other DNA binding proteins. For reasons that are not understood, a transcriptional activator can turn on transcription of a gene when bound many hundreds of base pairs from the gene in higher eukaryotes, but in yeast the activator must be bound much closer to the gene to work efficiently. We have modified yeast so as to allow 'activation at a distance', and we are now analyzing the mechanism to see whether the effect is consistent with the recruitment model. And we are examining the properties of a protein that seems to have a unique ability to trigger gene expression when artificially recruited to a gene. In particular we are testing the idea that this protein, a component of the large complex called the Mediator, is rapidly degraded in cells, and that this instability facilitates activation of transcription

描述（由申请人提供）：基因表达的调节是不同的生物学过程，包括从受精卵中开发生物体，生物对细胞外信号的反应以及记忆的内心。该赠款要求资金继续我们对基因调节机制的分析，这将有助于我们了解正常过程，并阐明它们如何出现。拟议的研究主要使用酵母细胞，这是一种真核生物，其中基因调节的许多方面与哺乳动物细胞的基因调节密切相关。在一系列论文和书籍中，我们开发了一个模型（称为“招聘”模型），用于转录调节器如何确定是否会激活或抑制任何给定的基因。我们在这里提出了一系列以各种方式探测和测试此模型的实验。我们已经开发了通过DNA结合激活剂募集的各种蛋白质复合物的特定基因外观的方法。必须将100多种蛋白质带到基因进行转录进行进行进行，我们正在使用我们的方法研究这种大复合物的逐步形成。在相关的发展中，我们现在可以精确地测定核小体对DNA的处理，并且我们正在研究核小体定位和去除如何受其他DNA结合蛋白的影响。出于尚不清楚的原因，当从较高的真核生物中绑定了数百个基因的基因对时，转录激活因子可以打开基因的转录，但是在酵母中，激活因子必须更接近基因才能有效地工作。我们已经修改了酵母，以允许“在距离处激活”，现在我们正在分析该机制，以查看效果是否与募集模型一致。我们正在研究蛋白质的特性，该蛋白质在人为地募集到基因时似乎具有触发基因表达的独特能力。特别是我们正在测试这种蛋白质是称为介体的大复合物的成分，在细胞中迅速降解，并且这种不稳定性促进了转录的激活