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: 8502698
• 负责人: ROBERT J FLETTERICK
• 金额: $ 107.31万
• 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/8502698
• 关键词: Biochemical; Biochemistry; Bioinformatics; Biological Assay; Biophysics; California; Cardiovascular Diseases; Cell Fate Control; Cells; Clinical; Complex; DNA; Data; Development; Embryo; Enhancers; Event; Gene Expression; Genetic Transcription; Goals; Health; Hormonal; Hospitals; Image; In Vitro; Individual; Institutes; Knowledge; Laboratories; Malignant Neoplasms; Methodist Church; Methods; Modeling; Molecular; Molecular Biology; Mutagenesis; Mutation; Participant; Pathogenesis; Physiological; Post-Translational Protein Processing; Principal Investigator; Protein Structure Initiative; Proteins; Regenerative Medicine; Research; Research Institute; Research Personnel; San Francisco; Signal Transduction; Site; Somatic Cell; Stem Cell Research; Stem cells; Structure; Transcriptional Regulation; Universities; Work; X-Ray Crystallography; anticancer research; base; design; embryonic stem cell; in vivo; induced pluripotent stem cell; mutant; pluripotency; programs; promoter; protein complex; research study; response; self-renewal; three dimensional structure; tool; transcription factor

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.

描述（由申请人提供）：该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形成和功能的分子机制。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，并分析它们的功能关联。将通过生物化学和生物物理方法评估数千个组件，以确定 X 射线晶体学目标的关键组件。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。它们中的每一个都代表了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进行深思熟虑的分析将使我们能够在这些部分图像之间建立适当的联系，并重建该转录机制关键参与者功能的通用模型。我们将证明该模型以及在使用 iPS 细胞的突变和功能研究中所观察到的已识别转录复合物内的调节相互作用的合理性。这些实验将在多个地点进行：卫理公会医院研究所（休斯顿）、加州大学旧金山分校生物化学和生物物理学系、格拉德斯通心血管疾病研究所（UCSF）以及 PSI 实验室的 X 射线晶体学。拟议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可供生化和临床研究人员使用，因此将对干细胞研究和再生医学产生重大影响。