Structures of Protein Complexes Regulating Transcription in Enbryonic Stem Cells

调节胚胎干细胞转录的蛋白质复合物的结构

• 批准号: 7982305
• 负责人: ROBERT J FLETTERICK
• 金额: $ 120.7万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-30 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7982305
• 关键词: Biochemical; Biochemistry; Bioinformatics; Biophysics; Cardiovascular Diseases; Cell Fate Control; Cells; Clinical; Complex; Data; Development; Disease; Drug Delivery Systems; Embryo; Genetic Transcription; Goals; Hospitals; Image; Institutes; Knowledge; Malignant Neoplasms; Methodist Church; Methods; Modeling; Molecular; Participant; Pathogenesis; Protein Structure Initiative; Regenerative Medicine; Research; Research Institute; Research Personnel; Site; Stem Cell Research; Stem cells; Structure; Tissues; X-Ray Crystallography; anticancer research; design; embryonic stem cell; induced pluripotent stem cell; novel; pluripotency; programs; protein complex; public health relevance; research study; self-renewal; three dimensional structure; tool

DESCRIPTION (provided by applicant): 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 interactions 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 of the 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. PUBLIC HEALTH RELEVANCE: The proposed studies on embryonic and reprogrammed stem cells will reveal parts of the complex mechanisms controlling cell fate, including those underlying self renewal, differentiation, and development of cancer. The results of our research will provide more efficient molecular tools allowing precise control over cell programming and reprogramming. The images and concepts that we produce will have immediate impact on regenerative medicine as well as stem cell and cancer research.

描述（由申请人提供）：本提案的主要目标是揭示胚胎干（ES）细胞和诱导多能性细胞（诱导多能干（iPS）细胞）所必需的关键转录组件的形成和功能的分子机制。利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，并分析它们的功能关联。成千上万的组件将通过生物化学和生物物理方法进行评估，以确定X射线晶体学的关键目标。我们的目标是确定 约100个稳定的多组分转录组装体的三维结构。它们中的每一个都将代表控制多能细胞特定转录景观的复杂转录机制的部分图像。我们期望对这些结构的深入分析将使我们能够在这些部分图像之间建立适当的联系，并重建这种转录机制的关键参与者的功能的一般模型。我们将证明这一模型和观察到的监管内确定的转录复合物的突变和功能研究，使用iPS细胞的相互作用。实验将在多个地点进行：卫理公会医院研究所（休斯顿），UCSF生物化学和生物物理系，Gladstone心血管疾病研究所（UCSF）和PSI实验室的X射线晶体学。这些结构和功能的研究将推动我们对控制细胞命运的基本机制的全面了解，包括那些潜在的自我更新，分化和癌症发病机制。这项研究的结果还将提供更有效的分子工具，允许精确控制细胞编程和重编程。积累的结构和功能数据将立即提供给生物化学和临床研究人员，因此，将对干细胞研究和再生医学产生重大影响。 公共卫生关系：对胚胎干细胞和重编程干细胞的拟议研究将揭示控制细胞命运的复杂机制的一部分，包括那些潜在的自我更新，分化和癌症发展。我们的研究结果将提供更有效的分子工具，允许精确控制细胞编程和重编程。我们产生的图像和概念将对再生医学以及干细胞和癌症研究产生直接影响。