Mechanistic studies of corepressor-mediated PPARγ transcriptional repression

辅阻遏物介导的 PPARγ 转录抑制的机制研究

• 批准号: 10591718
• 负责人: Douglas Kojetin
• 金额: $ 52.76万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-03-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10591718
• 关键词: Affect; Affinity; Agonist; Binding; Biochemical; Biological Assay; Biology; Cell model; Cell physiology; Cells; Chromatin; Complement; Coupled; Crystallization; Data; Electron Spin Resonance Spectroscopy; Gene Expression; Genes; Genetic Transcription; Goals; Insulin Resistance; Knowledge; Length; Ligand Binding Domain; Ligands; Link; Mediating; Methods; Molecular; Molecular Conformation; Molecular and Cellular Biology; Mutagenesis; NMR Spectroscopy; Nuclear Receptors; Obesity; Outcome; PPAR gamma; Peptides; Pharmaceutical Chemistry; Pharmacology; Phosphorylation; Proteins; Publishing; Repression; Structure; Work; X-Ray Crystallography; analog; antagonist; base; design; gene repression; genetic corepressor; insulin sensitizing drugs; lipid biosynthesis; promoter; recruit; scaffold; small molecule; structural biology; transcription factor; transcriptome; transcriptome sequencing

Peroxisome proliferator-activated receptor gamma (PPARγ) is a nuclear receptor transcription factor that regulates cellular differentiation, adipogenesis, and insulin resistance by recruiting transcriptional coregulator proteins (corepressor and coactivator proteins) to target gene promoters in a ligand-dependent manner. Structure-function approaches have defined how the transcriptionally active structural conformation and coactivator-selective functions of PPARγ are influenced by agonist ligands. However, little is known about the transcriptionally repressive conformation and corepressor-selective functions of PPARγ. Our long-term goal is to close this knowledge gap by defining how different pharmacological PPARγ ligands influence the structure and function of PPARγ between transcriptionally active and repressive states. In preliminary studies, we solved crystal structures of the PPARγ ligand-binding domain (LBD) in a transcriptionally repressive state using a unique corepressor-selective ligand, revealing a unique structural conformation that we have started to validate using solution NMR methods. In this project, we will use mechanistic studies to define how small molecule ligands impact PPARγ activation and repression on the molecular, structural, and cellular levels. Successful outcomes from our studies will define the activity-dependent conformational ensemble of the PPARγ LBD, develop ligands with enhanced corepressor-selective activity, and determine the molecular mechanisms by which corepressor-selective repressive ligands modulate PPARγ cellular functions.

过氧化物酶体增殖体激活受体γ (PPARγ)是一种核受体转录因子