The TFIID Complex-Biophysical & Structural Studies

TFIID 复合体-生物物理

• 批准号: 6840405
• 负责人: RAYMOND H JACOBSON
• 金额: $ 28.2万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-01-01 至 2008-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6840405
• 关键词: DNA directed RNA polymerase; Saccharomyces cerevisiae; analytical ultracentrifugation; calorimetry; crystallization; electron microscopy; enzyme activity; gel filtration chromatography; gene expression; genetic mapping; genetic transcription; histones; posttranslational modifications; protein binding; protein protein interaction; protein sequence; protein structure; transcription factor

DESCRIPTION (provided by applicant): In eukaryotic organisms, transcription of the messages contained within the genome is a tightly regulated process. Transcription occurs via the coordinated action of some 40-50 distinct protein components assembled into multiple complexes that cooperate to recognize transcription start sites, perform integration of regulatory signals, and provide the enzymatic activities necessary for the synthesis of RNA transcripts. Present within each gene are control elements that dictate how and when it is transcribed. For protein-coding genes transcribed by RNA polymerase II, interactions between the core promoter and the basal machinery nucleate formation of the pre-initiation complex - an assembly containing RNA Pol II and the general transcription factors, IIA, lIB, lID, lIE, IIF and IIH, while the regulatory elements are the targets of sequence-specific DNA-binding proteins that contain domains that affect activation or repression of transcription. The cumulative effects of gene specific activators/repressors interacting with regulatory sequence elements, as well as interactions between the core promoter elements and basal machinery, dictate the strength of transcription from any particular gene. The importance of the correct regulation of gene expression cannot be overstated. Errors in transcriptional control may lead to numerous deleterious outcomes that include but are not limited to, developmental defects, uncontrolled cell proliferation, inappropriate apoptosis, and metabolic diseases. An essential component for the proper regulation of transcription is general transcription factor lID (TFIID). It is a multi-subunit complex comprised of the TATA binding protein (TBP) and 13-15 additional subunits that not only nucleates assembly of the PIC, but is also the target and transducer of numerous transcriptional regulators. We wish to understand how the multiple components of the TFIID complex work together to facilitate precisely regulated gene expression which is essential for human health. To this end we will biochemically analyze and pursue high-resolution structural studies of TFIID or its constituent subunits. We will examine interactions between TFIID and activators or chromatin and characterize intrinsic enzymatic activities present within the TFIID complex. Using several complementary biophysical techniques, we aim to gain a detailed understanding of the structure/function relationships for this central component in the RNA Pol II transcription complex.

描述（由申请人提供）：在真核生物中，基因组中包含的消息的转录是一个严格调节的过程。转录通过约40-50种不同蛋白质成分的协调作用，这些蛋白质成分组装成多个复合物，这些络合物合作以识别转录起始位点，进行调节信号的整合，并提供合成RNA转录本所需的酶活性。每个基因中存在的是控制元素如何和​​何时转录的元素。对于由RNA聚合酶II转录的蛋白质编码基因，核心启动子与基础机械之间的相互作用与原始络合物的基础机械形成形成 - 一种包含RNA POL II的组装和一般的转录因子以及IIA，LIB，LIB，LID，LID，LID，IIF，IIF，IIF，IIF IIH，而调节元件是序列特异性DNA结合蛋白的靶标，该蛋白包含影响转录激活或抑制的域。基因特异性激活剂/阻遏物与调节序列元件相互作用的累积效应，以及核心启动子元素和基础机械之间的相互作用，决定了任何特定基因的转录强度。正确调节基因表达的重要性不能被夸大。转录控制中的错误可能导致许多有害结果，包括但不限于发育缺陷，不受控制的细胞增殖，不适当的凋亡和代谢疾病。适当调控转录的基本成分是一般转录因子盖（TFIID）。它是由TATA结合蛋白（TBP）和13-15个附加亚基组成的多生成复合物，不仅是PIC的组装，而且是众多转录调节剂的目标和换能器。我们希望了解TFIID复合物的多个组成部分如何共同起作用，以促进对人类健康至关重要的精确调节基因表达。为此，我们将生化分析和进行TFIID或其组成亚基的高分辨率结构研究。我们将检查TFIID与活化剂或染色质之间的相互作用，并表征TFIID复合物中存在的内在酶促活性。使用几种互补的生物物理技术，我们旨在详细了解RNA Pol II转录复合物中该中心成分的结构/功能关系。