Structural Characterization of Metabolic Gene Regulators

代谢基因调控因子的结构表征

• 批准号: 8339923
• 负责人: FRAYDOON RASTINEJAD
• 金额: $ 42.41万
• 依托单位: SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-29 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8339923
• 关键词: Acetylation; Affect; Architecture; Arginine; Binding; Biochemical; Cells; Complex; Coupled; DNA; DNA Binding; DNA Binding Domain; Deuterium; Diabetes Mellitus; Dimerization; Gene Activation; Genes; Glucose; Goals; Hydrogen; Insulin; Islet Cell; Islets of Langerhans; Learning; Ligand Binding; Link; Liver; Maps; Mass Spectrum Analysis; Metabolic; Methylation; Modification; Mutation; Nuclear Receptors; Pancreas; Peptides; Phosphorylation; Post-Translational Protein Processing; Property; Proteins; Regulation; Regulator Genes; Response Elements; Role; Series; Serine; Structure; Surface; Techniques; Testing; Thermodynamics; Time; Ubiquitination; Validation; X-Ray Crystallography; base; dimer; glucose production; human HNF4A protein; novel; programs; receptor

DESCRIPTION (provided by applicant): This proposal focuses on the structural and functional characterization of the hepatocyte nuclear factor 4 alpha (HNF4a), a master transcriptional regulator of metabolic gene programs. HNF4a does not respond to ligand binding, but instead appears to rely on methylation and phosphorylation, and on the induction of coactivators for regulating its transcriptional activity. Our recent structural characterization of HNF4a reveals multiple domain-domain interactions that were unanticipated. A highly interconnected domain arrangement links subunit dimerization to both DNA and coactivator binding. We wish to validate this understanding of the receptor's allosteric arrangement using a series of biochemical and cell-based studies that probe these inter-domain connections. We also found several posttranslational modifications that map to key domain-domain junctions. Therefore, a second goal is to understand the allosteric mechanism by which covalent modifications are transmitted to the DNA binding domain of the receptor. The third goal is to expand our structural and mechanistic understanding of how coactivators interact with HNF4a. Here we will test a novel hypothesis that some coactivators, including PGC-1a, can physically drive and stabilize the productive complex of HNF4a on DNA.

描述（由申请人提供）：该提案的重点是代谢基因程序的主要转录调节剂肝细胞核因子4α（HNF4A）的结构和功能表征。 HNF4A对配体结合反应，而是依赖于甲基化和磷酸化，以及诱导共激活因子来调节其转录活性。我们最近对HNF4A的结构表征揭示了多个域域相互作用，这些相互作用是意外的。高度互连的域布置将亚基二聚化与DNA和共激活因子结合。我们希望使用一系列探测这些域间连接的生化和基于细胞的研究来验证对受体变构的理解。我们还发现了几种映射到关键域域连接处的翻译后修改。因此，第二个目标是了解将共价修饰传递到受体的DNA结合结构域的变构机制。第三个目标是扩展我们对共激活因子如何与HNF4A相互作用的结构和机械理解。在这里，我们将检验一个新的假设，即某些共同激活因子（包括PGC-1A）可以在DNA上物理驱动和稳定HNF4A的生产络合物。