ARCHITECTURE OF THE EUKARYOTIC TRANSCRIPTION APPARATUS IN SOLUTION

解决方案中真核转录装置的架构

• 批准号: 8170044
• 负责人: David A. Bushnell
• 金额: $ 0.34万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8170044
• 关键词: Architecture; Complex; Computer Retrieval of Information on Scientific Projects Database; Cryoelectron Microscopy; Crystallography; Development; Electron Microscopy; Enzymes; Eukaryota; Funding; Future; Gene Expression; General Transcription Factors; Genetic Transcription; Grant; Institution; Mediator of activation protein; Messenger RNA; Pathway interactions; Process; RNA Polymerase II; Regulation; Research; Research Personnel; Resources; Solutions; Source; Structure; Transcription Repressor/Corepressor; Transcriptional Regulation; United States National Institutes of Health; base; dalton; meetings; polypeptide; promoter; tumorigenesis

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Transcription is the first step and the key control point in the pathway of gene expression. Transcriptional regulation underlies development, oncogenesis, and other fundamental processes. In eukaryotes the enzyme RNA Polymerase II (pol) is responsible for transcription of messenger RNA making pol?s regulation central to gene expression. Pol contains 12 subunits massing about 0.5 Mda, but despite its size and complexity pol cannot recognize and initiate transcription from a specific promoter. In order to transcribe from a specific promoter pol requires at least five general transcriptions factors (IID, IIB, IIE, IIF and IIH). In addition to the five general transcription factors, pol requires the action of a 1.5 MDa complex called mediator in order to respond to activators and repressors of transcription. The purpose of this proposal is to use solution scattering to study the various complexes involved in activated transcription up to and including the 60 polypeptide complex of pol, the 5 general transcription factors and mediator. Structure determination of these large assemblies presents formidable technical challenges. We are attempting to meet these challenges through a combination of x-ray crystallography, electron microscopy, and solution x-ray scattering. This experimental proposal constitutes our research effort using solution scattering, which we believe fills the technological gap between x-ray crystallography and cryoEM in terms of its strength on submega Dalton complexes and its ability to study them in solution. The significance of the proposed research may be summarized as follows: it will provide the structural information needed to fully understand the fundamental mechanism of transcription; it will establish a structural basis for studies of transcriptional regulation; and it may indicate how structures of other heterogeneous multiprotein assemblies can be solved in the future by the combined structural approach.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 转录是基因表达途径中的第一步和关键控制点。 转录调节是发展，​​肿瘤发生和其他基本过程的基础。 在真核生物中，酶RNA聚合酶II（POL）负责使Messenger RNA的转录使POL的调节与基因表达中心。 POL包含12个亚基量约为0.5 MDA，但是尽管其大小和复杂性，POL无法识别并启动特定启动子的转录。 为了从特定的启动子转录，Pol需要至少五个一般转录因子（IID，IIB，IIE，IIF和IIH）。 除了五个一般的转录因子外，POL还需要1.5 MDA复合物的作用，称为介体，以响应转录的激活剂和阻遏物。该提案的目的是使用溶液散射来研究激活转录涉及的各种复合物，直至和包括60个POL的60多肽复合物，这是5个常规转录因子和介体。这些大型组合的结构确定提出了巨大的技术挑战。我们试图通过X射线晶体学，电子显微镜和溶液X射线散射的结合来应对这些挑战。这项实验性建议构成了我们使用溶液散射的研究工作，我们认为这可以填补X射线晶体学和冷冻仪之间的技术差距，从而在其对subsega dalton综合体的强度以及在解决方案中研究它们的能力。拟议研究的重要性可以总结如下：它将提供充分了解转录基本机制所需的结构信息；它将建立转录调节研究的结构基础；它可能表明如何通过组合结构方法在将来解决其他异质多蛋白组件的结构。