Structural studies of the eukaryotic transcription

真核转录的结构研究

• 批准号: 6520489
• 负责人: Eva Nogales
• 金额: $ 12.03万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-05-01 至 2006-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6520489
• 关键词: DNA binding protein; DNA directed RNA polymerase; atomic force microscopy; conformation; cryoelectron microscopy; eukaryote; genetic transcription; nucleic acid structure; protein binding; structural biology; transcription factor

DESCRIPTION (provided by applicant): Regulation of gene transcription is essential in the process of cell differentiation and development of the organism, as well as for the survival of each cell in changing environmental conditions. The importance of gene transcription and its regulation is underlined by the myriad of human disorders that result from the malfunction of the molecular components involved in this cellular process. Initiation of gene transcription in eukaryotes starts with the binding of TFIID, containing the TATA binding protein (TBP), to the DNA core promoter, and the consequent assembly of the general transcriptional machinery. Six general transcription factors and RNA Pol H constitute the minimal protein assembly for transcription, adding up to several million Daltons. TFIID is also involved in the interaction with gene-specific activators. Our ultimate goal is to understand how the eukaryotic transcription initiation machinery assembles and functions, and the structural mechanisms for gene transcription regulation. To this end we will characterize the structure of TFIID and its interaction with other general factors and with activators using cryo-electron microscopy and single-particle image reconstruction. We have recently obtained an initial structure of TFHD and its complex with full length TFILA and TFIIB at 35 A resolution by negative stain, and localized, by antibody mapping, the position of TBP within the complex. One of our immediate priorities is to obtain a 3-D reconstruction of frozen-hydrated human TFIID complexes at a resolution better than 25 A. We are interested in following the structural assembly and conformational changes of this complex machinery by generating medium resolution structures of increasingly larger complexes, with TFIID as the central core. Our final goal is to characterize the structural basis of transcription regulation by the interaction of the general transcriptional machinery with activators. To this end we will study the complexes formed by TFIID with the general activator Sp 1.

描述（由申请人提供）：基因转录的调节为 在细胞分化和发展过程中至关重要的 有机体以及每个细胞在改变环境中的生存 状况。基因转录及其调节的重要性是 由由于故障的无数人类疾病而强调 参与此细胞过程的分子成分。基因的启动 真核生物中的转录始于tfiid的结合，包含 TATA结合蛋白（TBP），与DNA核心启动子，结果 一般转录机械的组装。六个一般转录 因素和RNA pol H构成最小蛋白质组件 转录，总计多达数百万美元。 TFIID也参与了 与基因特异性激活剂的相互作用。我们的最终目标是 了解真核转录启动机械如何组装和 功能和基因转录调控的结构机制。到 我们将表征TFIID的结构及其与之相互作用 其他一般因素以及使用冷冻电子显微镜和 单粒子图像重建。我们最近获得了最初的 TFHD的结构及其复合物，具有全长Tfila和TFIIB在35 a处的结构 通过负污渍和局部通过抗体映射的局部分辨率，该位置 综合体内的TBP。我们的直接优先事项之一是获得3-D 在分辨率更好地重建冷冻水合的人类TFIID复合物 超过25 A.我们有兴趣遵循结构组装和 通过生成培养基的构象变化 越来越大的复合物的分辨率结构，TFIID为 中心核心。我们的最终目标是表征 转录调节通过一般转录的相互作用 带有激活器的机械。为此，我们将研究由 带有通用激活剂SP 1的TFIID。