Structure and Mechanism of Eukaryotic Transcription Regulation

真核生物转录调控的结构和机制

基本信息

批准号：
10625407
负责人：
Yuan He
金额：
$ 35.34万
依托单位：
NORTHWESTERN UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-06-01 至 2026-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10625407
关键词：
ATP Hydrolysis Address Architecture BRF1 gene Binding Biochemical Biogenesis Cell Nucleus Cells Chemicals Chromosomes Code Complex Core Assembly Cryoelectron Microscopy DNA DNA-Directed RNA Polymerase Data Elements Enhancers Enzymes Eukaryota GTF2IRD1 gene Gene Expression Gene Expression Regulation Gene Proteins Genes Genetic Genetic Transcription Goals Human In Vitro Interferon-beta Mass Spectrum Analysis Mediator Molecular Molecular Biology Organism Polymerase Process Production Protein Biosynthesis Proteins RNA RNA Polymerase II RNA chemical synthesis RNA, Ribosomal, 5S RNA, ribosomal, 25S Recycling Resolution Ribosomal RNA Series Signal Pathway Signal Transduction Structure System TAF1 gene Techniques Time Transcription Factor TFIIA Transcription Factor TFIIB Transcription Factor TFIIIA Transcription Factor TFIIIB Transcription Initiation Transcription Initiation Site Transcriptional Activation Transcriptional Regulation Transfer RNA Untranslated RNA Visualization Yeasts cell growth crosslink genetic information human disease interdisciplinary approach novel particle polypeptide promoter rRNA Genes reconstitution recruit transcription factor transcription factor TFIIE transcription factor TFIIF transcription factor TFIIH transcription factor TFIIIC translocase

项目摘要

Summary Transcription is the first step to read out the genetic information stored in the chromosome. In eukaryotes, three multi-subunit RNA polymerases (Pols), Pol I, II, and III transcribe the 25S ribosomal RNA (rRNA), protein-coding, and short non-coding RNAs such as transfer RNA (tRNA) and 25S rRNA genes, respectively. A wealth of genetic and biochemical data accumulated over 50 years have uncovered most of the molecular players involved and details of the intricate regulatory circuits. The recruitment and assembly of the initiation complex at the promoter represent one of the key regulatory steps during gene expression. The Pol II initiation machinery includes the activator-bound Mediator along with a series of general transcription factors (GTFs) (TFIID, TFIIA, TFIIB, TFIIE, TFIIF, and TFIIH) that assemble into a ~4-megadalton (MDa) pre-initiation complex (PIC) on core promoter DNA. On the other hand, the Pol III enzyme is recruited onto its promoter with the help of corresponding GTFs (TFIIIC, TBP, BRF1, and B''). This proposal aims to investigate the mechanism of transcription regulation by directly visualizing these transcriptional competent initiation complexes using single-particle cryo-EM. Comparable pictures of these different but evolutionarily related systems at this critical step of gene regulation will provide an unprecedented, comprehensive view of this key process in the central dogma of molecular biology.

概括转录是读出染色体中存储的遗传信息的第一步。在真核生物中，三种多亚基 RNA 聚合酶 (Pols)，Pol I、II 和 III 转录 25S 核糖体 RNA (rRNA)，蛋白质编码RNA和短非编码RNA，例如转移RNA (tRNA)和25S rRNA基因。一个 50多年来积累的丰富的遗传和生化数据已经揭示了大部分分子涉及的参与者以及复杂的监管电路的细节。入会的招募和集合启动子处的复合物代表基因表达过程中的关键调控步骤之一。 Pol II 启动机制包括激活剂结合的介体以及一系列通用转录因子 (GTF) （TFIID、TFIIA、TFIB、TFIIE、TFIIF 和 TFIIH）组装成约 4 兆道尔顿 (MDa) 预引发核心启动子DNA上的复合物（PIC）。另一方面，Pol III 酶被招募到其启动子上相应 GTF（TFIIIC、TBP、BRF1 和 B''）的帮助。本提案旨在调查通过直接可视化这些转录活性起始来了解转录调控机制使用单粒子冷冻电镜形成复合物。这些不同但进化相关的可比图片基因调控这一关键步骤的系统将为这一关键步骤提供前所未有的全面视图分子生物学中心法则的过程。