RETROVIRAL TRANSCRIPTIONAL CONTROL MECHANISMS

逆转录病毒转录控制机制

• 批准号: 6635959
• 负责人: Barbara J Graves
• 金额: $ 31.5万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-07-01 至 2005-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6635959
• 关键词: Caenorhabditis elegans; DNA binding protein; biological signal transduction; calcium ion; conformation; gene induction /repression; genetic promoter element; nuclear magnetic resonance spectroscopy; phosphorylation; protein protein interaction; site directed mutagenesis; transcription factor; yeast two hybrid system

This proposal investigates the regulatory transcription factor Ets-1 as a model system for understanding the pathways to specificity within a gene family of DNA binding proteins. Ets-1 belongs to the ets family of transcription factors whose members share common DNA binding proteins. Ets-1 belong to the ets family of transcription factors whose members share common DNA binding properties. Our approaches are distinctive due to the extensive structural framework we have developed for Ets-1 and the rigorous quantitative analyses that we routinely perform. A major focus is the auto-inhibition phenomenon of Ets-1 in which DNA binding of the ETS domain is repressed by an intramolecular mechanism. Our recent experiments identified two sources of regulation that appear to act through the auto-inhibition mechanism. This proposal investigates both sources of regulation. 1. First, we will determine the mechanism by which Ca2+-dependent phosphorylation reinforces auto-inhibition, thus further repressing DNA binding. 2. A partnership between CBFalpha2 and Ets-1 counteracts the effects of auto-inhibition. Our goal is to test a model of this mechanism by obtaining structural data for the Ets-1: CBF ternary complex. We will also search for in vivo evidence for this partnership using chromatin immunoprecipitations. 3. The third specific aim focuses on the Pointed (PNT) domain of Ets-1. This region shows conservation within the ets family; however, a variety of functions have been proposed, including interactions with co-activators, co-repressors and self-association. We plan to directly test the oligomerization hypothesis and screen for interacting proteins. 4. Finally, in the fourth aim we develop a genetic approach to investigate the ets gene family within the C. elegans system. The 10 ets genes in C. elegans will be surveyed both for expression and by analysis of null phenotypes. Our initial focus is the function of the PNT domain within two C. elegans ets genes. Our initial focus is the function of the PNT domain within two C. elegans ets genes. Our work on the ets gene family is relevant to human disease. Ets-1 regulates gene expression of the Moloney murine leukemia virus and HIV-1, and we use transcriptional control elements from these viruses as model enhancers in our experiments. Second, the ets proteins, specifically the ETS and PNT domains, are involved in human cancers caused by chromosome translocations.

该提案研究了调节转录因子ETS-1作为一种模型系统，用于理解DNA结合蛋白基因家族中特异性的途径。 ETS-1属于转录因子的ETS家族，其成员具有共同的DNA结合蛋白。 ETS-1属于转录因子的ETS家族，其成员具有共同的DNA结合特性。由于我们为ETS-1开发了广泛的结构框架以及我们通常执行的严格定量分析，因此我们的方法是独特的。一个主要重点是ETS-1的自身抑制现象，其中ETS结构域的DNA结合被分子内机制抑制。我们最近的实验确定了似乎通过自动抑制机制起作用的两个调节来源。该提案调查了两个法规的来源。 1。首先，我们将确定Ca2+依赖性磷酸化增强自身抑制的机制，从而进一步抑制DNA结合。 2。CBFALPHA2和ETS-1之间的伙伴关系抵消了自动抑制的影响。我们的目标是通过获得ETS-1：CBF三元复合物的结构数据来测试该机制的模型。我们还将使用染色质免疫沉淀搜索这种伙伴关系的体内证据。 3。第三个特定目的侧重于ETS-1的尖头（PNT）域。该地区显示了ETS家族中的保护；但是，已经提出了多种功能，包括与共激活因子，共抑制器和自我关联的相互作用。我们计划直接测试相互作用蛋白的寡聚假说和筛选。 4。最后，在第四个目标中，我们开发了一种遗传方法来研究秀丽隐杆线虫系统中的ETS基因家族。秀丽隐杆线虫中的10个ETS基因将通过表达和分析无效表型进行调查。我们最初的焦点是两个秀丽隐杆线虫ETS基因中PNT结构域的功能。我们最初的焦点是两个秀丽隐杆线虫ETS基因中PNT结构域的功能。我们在ETS基因家族方面的工作与人类疾病有关。 ETS-1调节Moloney鼠白血病病毒和HIV-1的基因表达，我们将这些病毒中的转录控制元件用作实验中的模型增强子。其次，ETS蛋白，特别是ETS和PNT结构域，参与由染色体易位引起的人类癌。