Consortia for High-Throughput-Enabled Structural Biology Partnerships (U01)

高通量结构生物学合作联盟 (U01)

• 批准号: 8379000
• 负责人: Paul Webb
• 金额: $ 17.79万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8379000
• 关键词: Consortia; Throughput; Enabled; Structural; Biology

The major goal of this proposal is to reveal molecular mechanisms underlying formation and function of critical transcriptional assemblies essential to embryonic stem (ES) cells and cells with induced pluripotency (induced pluripotent stem (iPS) cells). Using bioinformatics and high throughput experimental methods, we will prepare defined domains of critical transcriptional factors controlling cell pluripotency and analyze them in their functional associations. Thousands of assemblies will be evaluated by biochemical and biophysical methods to identify the critical ones to be targeted by X-ray crystallography. We aim for determination of three-dimensional structures for about 100 stable multi-component transcriptional assemblies. Each of them will represent a partial image of complicated transcriptional machinery controlling the specific transcriptional landscape of pluripotent cells. We expect that thoughtful analyses of these structures will enable us to establish the proper connections between these partial images and reconstruct a general model for function of critical participants of this transcriptional machinery. We will justify this model and the observed regulatory nteractions within identified transcriptional complexes in mutational and functional studies using iPS cells. The experiments are to be done at multiple sites: The Methodist Hospital Research Institute (Houston), Department of Biochemistry and Biophysics at UCSF, the Gladstone Institute of Cardiovascular Disease (UCSF) and X-ray crystallography by the PSI labs. The proposed structural and functional studies will propel our general knowledge ofthe basic mechanisms controlling cell fate, including those underlying self renewal, differentiation and pathogenesis of cancer. The results of this research will also provide more efficient molecular tools allowing precise control over cell programming and reprogramming. The accumulated structural and functional data would be immediately available to biochemical and clinical researchers, and therefore, would have a major impact on stem cell research as well as regenerative medicine.

该提案的主要目的是揭示形成和功能的分子机制 具有诱导多能性的胚胎干细胞（ES）细胞和细胞必不可少的关键转录组件 （诱导多能茎（IPS）细胞）。使用生物信息学和高通量实验方法，我们 将准备控制细胞多能性的关键转录因子的定义域并分析它们 在其功能关联中。将通过生化和生物物理评估成千上万的组件 识别X射线晶体学针对的关键方法的方法。我们的目标是确定 大约100个稳定的多组分转录组件的三维结构。他们每个人 将代表控制特定转录的复杂转录机械的部分图像 多能细胞的景观。我们希望对这些结构进行周到的分析将使我们能够 在这些部分图像之间建立适当的连接并重建功能的通用模型 该转录机械的关键参与者。我们将证明这一模型和观察到的调节 使用IPS细胞的突变和功能研究中已鉴定的转录复合物中的Nteractions。 实验将在多个地点进行：卫理公会医院研究所（休斯顿）， 格拉德斯通心血管疾病研究所UCSF生物化学和生物物理学系 PSI实验室（UCSF）和X射线晶体学。提出的结构和功能研究将推动 我们对控制细胞命运的基本机制的一般知识，包括基本的自我更新， 癌症的分化和发病机理。这项研究的结果还将提供更有效的效率 分子工具，允许精确控制细胞编程和重编程。积累 结构和功能数据将立即用于生化和临床研究人员，以及 因此，将对干细胞研究和再生医学产生重大影响。