New Paradigms for the molecular basis of RNA polymerase I transcription

RNA 聚合酶 I 转录分子基础的新范式

• 批准号: 10178706
• 负责人: Bruce Alan Knutson
• 金额: $ 34.02万
• 依托单位: UPSTATE MEDICAL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-03-05 至 2026-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10178706
• 关键词: Address; Architecture; Binding Proteins; Biochemical; Biochemical Genetics; Biochemistry; Bioinformatics; Biological Assay; Biophysics; Cancer Etiology; Cell Proliferation; Cells; Complement; Complex; DNA; DNA Polymerase II; DNA Polymerase III; DNA-Protein Interaction; Defect; Diagnosis; Disease; Down-Regulation; Fibrinogen; Gene Expression Regulation; Genes; Genetic; Genetic Diseases; Genetic Transcription; Genomics; Goals; Health; Histones; Human; Hybrids; Intervention; Knowledge; Laboratory Study; Lead; Link; Location; Malignant Neoplasms; Maps; Methodology; Methods; Modeling; Molecular; Molecular Genetics; Molecular Structure; Mutation; Nutrient; Outcome; Outcome Study; Peptide Initiation Factors; Play; Polymerase; Positioning Attribute; Process; Proteins; Proteomics; RNA; RNA Polymerase I; Recombinant DNA; Regulation; Research; Ribosomal RNA; Ribosomes; Role; Signal Transduction; Stress; Structural Models; Structure; System; TATA Box; TATA-Box Binding Protein; Techniques; Testing; Therapeutic; Transcription Process; Transcriptional Activation; Translating; United States National Institutes of Health; Up-Regulation; Work; Yeasts; activating transcription factor; cell growth; craniofacial disorder; developmental disease; extracellular; human disease; innovation; interdisciplinary approach; new therapeutic target; novel; organizational structure; protein function; rRNA Precursor; reconstitution; structural biology; therapy development; transcription factor

Project Summary Eukaryotic RNA polymerase I (Pol I) transcribes ribosomal RNA, a key component of ribosomes. Pol I transcription accounts for the majority of the total RNA in cells, and its upregulation in human cells is a hallmark of cancer while its downregulation is a hallmark of several developmental disorders. Pol I transcription is understudied compared to transcription by Pol II and even Pol III. Our preliminary work suggests fundamental differences between Pol I and Pols II and III that are the basis for this proposal. Our broad long-term objectives are to determine the molecular mechanism of Pol I transcription and how its dysregulation leads to cancer and developmental disorders. There are major gaps in our understanding of (1) the structural organization and architecture of Pol I transcription complexes; (2) the mechanism for how Pol I initiation factors interact with rDNA, which encodes ribosomal RNA; and (3) the molecular function of several key Pol I transcription factors in the activation process. The first rationale for this work is that determining the mechanism and regulation of Pol I transcription will form the molecular basis for understanding how Pol I defects lead to human disease. Our central hypothesis is that Pol I factors use a unique mechanism to carry out transcription and their structure and function is different from the mechanisms governing Pol II and III transcription. The second rationale is that understanding the Pol I transcription mechanism at the most basic and fundamental levels will translate to a better understanding of the connection between Pol I and cancer, leading to new cancer therapeutic strategies. Our proposed research will use a conceptually and technically innovative cross-organismal and interdisciplinary approach that employs a combination of bioinformatic, computational, molecular, biochemical, genetic, genomic, proteomic, and structural methods in the yeast and human cells. Guided by strong preliminary studies, we will test two specific aims: (1) Determine the unique “coactivator” role of TATA-binding protein (TBP) in Pol I transcription, and (2) Determine the mechanism of Pol I transcription activation. To accomplish these aims, we will use well-established and complementary approaches to identify and map novel Pol I interactions in their native context. We will complement these studies with structural modeling in combination with molecular, genetic, and biochemical functional assays to identify Pol I factor functions conserved from yeast to humans. The proposed research is significant because it will lead to a detailed description of the Pol I transcription mechanism and will provide a conceptual framework for understanding the link between Pol I and human disease. Ultimately, this work will illuminate new avenues for diagnosis, potential interventions, and the development of therapies targeting these novel protein-protein and protein-DNA interactions.

项目概要 真核 RNA 聚合酶 I (Pol I) 转录核糖体 RNA，这是核糖体的关键成分。聚合酶 I 转录 占细胞总 RNA 的大部分，其在人类细胞中的上调是癌症的标志，而 它的下调是多种发育障碍的标志。 Pol I 转录尚未得到充分研究 到 Pol II 甚至 Pol III 的转录。我们的初步工作表明 Pol I 之间存在根本差异 以及作为本提案基础的 Pols II 和 III。我们广泛的长期目标是确定分子 Pol I 转录机制及其失调如何导致癌症和发育障碍。那里 我们对 (1) Pol I 转录的结构组织和架构的理解存在重大差距 复合体； (2) Pol I起始因子如何与编码核糖体RNA的rDNA相互作用的机制； (3)几个关键的Pol I转录因子在激活过程中的分子功能。第一个理由 这项工作的目的是确定 Pol I 转录的机制和调控将形成分子基础 了解 Pol I 缺陷如何导致人类疾病。我们的中心假设是 Pol I 因子使用 进行转录的独特机制，其结构和功能与机制不同 控制 Pol II 和 III 转录。第二个理由是了解 Pol I 转录机制 在最基本和根本的层面上将转化为更好地理解 Pol I 之间的联系 和癌症，从而产生新的癌症治疗策略。我们提出的研究将使用概念上和 技术创新的跨组织和跨学科方法，结合了生物信息学、 酵母和酵母中的计算、分子、生物化学、遗传、基因组、蛋白质组和结构方法 人体细胞。在强有力的初步研究的指导下，我们将测试两个具体目标：（1）确定独特的 TATA结合蛋白（TBP）在Pol I转录中的“共激活剂”作用，以及（2）确定Pol I的机制 转录激活。为了实现这些目标，我们将使用行之有效的补充方法 识别并绘制其原生环境中新颖的 Pol I 相互作用。我们将通过结构性研究来补充这些研究 建模与分子、遗传和生化功能分析相结合，以确定 Pol I 因子功能 从酵母到人类都是保守的。拟议的研究很重要，因为它将带来详细的描述 Pol I 转录机制的研究，并将提供一个概念框架来理解之间的联系 Pol I 和人类疾病。最终，这项工作将阐明诊断、潜在干预措施的新途径， 以及针对这些新型蛋白质-蛋白质和蛋白质-DNA 相互作用的疗法的开发。