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: 8533829
• 负责人: ROBERT J FLETTERICK
• 金额: $ 5.2万
• 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/8533829
• 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; 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

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.

本研究的主要目的是揭示细胞凋亡形成和功能的分子机制， 胚胎干（ES）细胞和具有诱导多能性的细胞所必需的关键转录组件 （诱导多能干（iPS）细胞）。利用生物信息学和高通量实验方法，我们 将制备控制细胞多能性的关键转录因子的定义域并分析它们 在他们的功能协会。数以千计的组件将通过生物化学和生物物理学进行评估。 方法，以确定关键的目标，由X射线晶体学。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。他们每个 将代表控制特定转录的复杂转录机制的部分图像， 多能细胞的景观。我们期望，对这些结构的深入分析将使我们能够 在这些局部图像之间建立适当的连接，并重建函数的通用模型 这个转录机制的关键参与者。我们将证明这个模型和观察到的监管 在使用iPS细胞的突变和功能研究中鉴定的转录复合物内的相互作用。 实验将在多个地点进行：卫理公会医院研究所（休斯顿）， 加州大学旧金山分校生物化学和生物物理学系，格莱斯顿心血管疾病研究所 （UCSF）和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一般知识，包括那些潜在的自我更新， 肿瘤的分化和发病机制。这项研究的结果也将提供更有效的 分子工具允许精确控制细胞编程和重编程。累计 生物化学和临床研究人员将立即获得结构和功能数据， 因此，将对干细胞研究以及再生医学产生重大影响。