The TFIID Complex-Biophysical & Structural Studies

TFIID 复合体-生物物理

• 批准号: 7163787
• 负责人: RAYMOND H JACOBSON
• 金额: $ 26.74万
• 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/7163787
• 关键词: Affect; Affinity; Affinity Chromatography; Antibodies; Apoptosis; Binding; Biochemical; Biochemical Genetics; Biological Assay; Bromodomain; Calorimetry; Categories; Cell Proliferation; Chromatin; Code; Complement Factor D; Complex; Crystallization; DNA; DNA Polymerase II; DNA Sequence; Data; Defect; Development; Electron Microscopy; Electrons; Elements; Endopeptidases; Fermentation; Gel Chromatography; Gene Expression; Gene Expression Regulation; General Transcription Factors; Genes; Genetic; Genetic Transcription; Genome; Health; Helix (Snails); Histone Code; Histone H3; Histone H4; Histones; Human; In Vitro; Kinetics; Lead; Libraries; Ligands; Localized; Lysine; Maps; Measures; Metabolic Diseases; Methods; Methylation; Modification; Molecular Machines; Negative Staining; Nucleosomes; Numbers; Organism; Outcome; Pattern; Peptide Hydrolases; Peptides; Phosphorylation; Play; Positioning Attribute; Post-Translational Protein Processing; Process; Production; Proteins; Proteolysis; Protocols documentation; RNA; RNA Polymerase II; RNA chemical synthesis; Recombinant Proteins; Recruitment Activity; Regulatory Element; Resolution; Role; Saccharomyces cerevisiae; Sedimentation process; Sequence-Specific DNA Binding Protein; Series; Set protein; Signal Transduction; Site; Specificity; Structure; Structure-Activity Relationship; System; T cell activating factor; TATA-Binding Protein Associated Factors; TATA-Box Binding Protein; Tail; Techniques; Titrations; Transcript; Transcription Factor TFIIA; Transcription Factor TFIIB; Transcription Initiation Site; Transcriptional Regulation; Transducers; Work; Yeasts; analytical ultracentrifugation; domain mapping; ear helix; expression vector; gene repression; mutant; programs; promoter; protein expression; seal; size; transcription factor; transcription factor TFIIE; transcription factor TFIIF; transcription factor TFIIH

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，基因组中包含的信息转录是一个严格调控的过程。转录是通过大约40-50种不同的蛋白质组分的协调作用而发生的，这些蛋白质组分组装成多个复合物，合作识别转录起始位点，执行调节信号的整合，并提供合成RNA转录物所需的酶活性。存在于每个基因内的控制元件决定了如何以及何时转录。对于RNA聚合酶II转录的蛋白质编码基因，核心启动子和基础机制之间的相互作用形成了起始前复合物-一个包含RNA Pol II和一般转录因子IIA， lIB, lID, lIE， IIF和IIH的组装体，而调控元件是序列特异性dna结合蛋白的靶标，这些蛋白含有影响转录激活或抑制的结构域。基因特异性激活因子/抑制因子与调控序列元件相互作用的累积效应，以及核心启动子元件与基础机制之间的相互作用，决定了任何特定基因的转录强度。正确调控基因表达的重要性再怎么强调也不为过。转录控制中的错误可能导致许多有害的结果，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。一般转录因子lID （TFIID）是正常调控转录的重要组成部分。它是一个多亚基复合物，由TATA结合蛋白（TBP）和13-15个额外的亚基组成，不仅是PIC组装的核，而且是许多转录调节因子的靶标和传感器。我们希望了解TFIID复合体的多个组成部分如何协同工作，以促进对人类健康至关重要的精确调控基因表达。为此，我们将对TFIID或其组成亚基进行生化分析和高分辨率结构研究。我们将研究TFIID与激活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活性。利用几种互补的生物物理技术，我们的目标是详细了解RNA Pol II转录复合体中这一核心成分的结构/功能关系。