A Global Characterization of the Interaction Between the AhR and KLF-6

AhR 和 KLF-6 之间相互作用的全局表征

• 批准号: 8785979
• 负责人: Daniel Jackson
• 金额: $ 3.06万
• 依托单位: UNIVERSITY OF TEXAS MED BR GALVESTON
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-01 至 2017-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8785979
• 关键词: AHR gene; Abbreviations; Amino Acids; Apoptosis; Aromatic Polycyclic Hydrocarbons; Aryl Hydrocarbon Receptor; Automobile Driving; BHLH Protein; Binding; Binding Sites; Biology; Cell Cycle; Cell Cycle Arrest; Cell Nucleus; Cell physiology; Co-Immunoprecipitations; Coin; Complex; Consensus; Coupled; Cytochrome P450; DNA; DNA Binding; Data; Dermal; Development; Diabetes Mellitus; Diagnostic; Dimethyl Sulfoxide; Dioxins; Disease; Electrophoretic Mobility Shift Assay; Elements; Environmental Pollution; Exposure to; Family; Functional disorder; Future; Gene Expression; Gene Expression Profile; Gene Expression Regulation; Gene Targeting; Genes; Genetic; Goals; Health; Hepatocyte; Hepatotoxicity; Herbicides; Heterodimerization; Human; Immune; Immunosuppression; In Vitro; Industrial Waste; Insulin; Knockout Mice; Knowledge; Learning; Ligands; Light; Liver; Liver diseases; Malignant Neoplasms; Meat; Mediating; Mediator of activation protein; Methods; Molecular; Molecular Chaperones; Mus; Mutagenesis; Mutation; Nuclear Translocation; Nucleotides; Oils; Oryctolagus cuniculus; Pathway interactions; Physiological Processes; Plasminogen Activator Inhibitor 1; Plasminogen Inactivators; Prevention; Process; Protein Binding; Proteins; RNA Sequences; Receptor Activation; Recombinants; Research; Response Elements; Reticulocytes; Role; Sequence Analysis; Shotgun Sequencing; Signal Transduction; Site; Smoke; System; Testing; Tetrachlorodibenzodioxin; Time; Toxic effect; Transcriptional Regulation; Tumor Suppressor Proteins; Weather; Work; Xenobiotics; activating transcription factor; aryl hydrocarbon receptor ligand; carcinogenesis; chromatin immunoprecipitation; detector; genome-wide; ground water; human ARNT protein; in vivo; inhibitor/antagonist; innovation; insight; next generation sequencing; novel; pollutant; prognostic; protein complex; protein protein interaction; public health relevance; receptor; receptor binding; research study; response; tool; transcription factor; transcriptome sequencing

DESCRIPTION (provided by applicant): The Aryl Hydrocarbon Receptor (AhR) is a basic helix-loop-helix transcription factor activated by diverse polycyclic aromatic hydrocarbons, including the prototypical ligand and persistent environmental contaminant 2,3,7,8-tetrachlorodibenzo-p-dioxin (dioxin,TCDD). Canonically, the ligand activated AhR translocates to the nucleus, dissociates from chaperones, and heterodimerizes with the Aryl Hydrocarbon Receptor Nuclear Translocator (Arnt) to transcriptionally regulate target genes via the cis DNA motif GCGTG, termed the xenobiotic response element (XRE). Recent work has shown a number of dioxin responsive AhR target genes lack an XRE. We have recently discovered the AhR to form a novel protein complex through an interaction with Kruppel-like Factor 6 (KLF6), which binds DNA via a novel non-consensus XRE (NC-XRE). We have subsequently shown that the KLF6/AhR complex regulates expression of a novel set of AhR genes, in response to dioxin, via binding to the NC-XRE. In this light, I hypothesize the KLF6/AhR heterodimer represents a novel regulatory complex conferring transcriptional control on novel target genes via binding to the NC-XRE. To investigate this novel protein interaction I will use in vivo chromatin immunoprecipitation next generation sequencing (ChIP-seq) experiments to identify the gene targets of this complex in the mouse liver coupled with whole transcriptome shotgun sequencing to anchor DNA binding of the KLF6/AhR transcription factor complex to changes in gene expression. This method will allow us to delineate functional vs. non-functional NC-XREs in KLF6/AhR target genes. I will examine the KLF6/AhR protein-protein interaction and identify the protein elements necessary for the interaction and characterize the key NC-XRE DNA motif elements necessary for KLF6/AhR binding. It is widely accepted that the AhR is required for the development of toxicity following exposure to dioxin. However, little is known about the mechanisms of the AhR driving the development of this toxicity. As our data suggest the NC-XRE represents a novel signaling cascade for the AhR following dioxin exposure, this work will provide novel insight on the mechanisms of dioxin toxicity with respect to the AhR, while shedding light on novel prognostic and diagnostics tools for identifying and treating the deleterious health effects associated with TCDD exposure.

描述（由申请人提供）：芳基烃受体（AhR）是一种基本的螺旋-环-螺旋转录因子，由多种多环芳烃激活，包括原型配体和持久性环境污染物2,3,7,8-四氯二苯并-p -二恶英（dioxin，TCDD）。通常，配体激活的 AhR 易位到细胞核，与伴侣分离，并与芳基烃受体核易位蛋白 (Arnt) 异二聚化，通过顺式 DNA 基序 GCGTG（称为异生素反应元件 (XRE)）转录调节靶基因。最近的工作表明，许多二恶英响应性 AhR 靶基因缺乏 XRE。我们最近发现 AhR 通过与 Kruppel 样因子 6 (KLF6) 相互作用形成一种新型蛋白质复合物，KLF6 通过新型非共有 XRE (NC-XRE) 结合 DNA。我们随后表明，KLF6/AhR 复合物通过与 NC-XRE 结合，调节一组新的 AhR 基因的表达，以响应二恶英。有鉴于此，我假设 KLF6/AhR 异二聚体代表一种新型调控复合物，通过与 NC-XRE 结合赋予新型靶基因转录控制。为了研究这种新型蛋白质相互作用，我将使用体内染色质免疫沉淀下一代测序 (ChIP-seq) 实验来鉴定小鼠肝脏中该复合物的基因靶标，并结合全转录组鸟枪法测序来锚定 KLF6/AhR 转录的 DNA 结合基因表达变化的复杂因素。该方法将使我们能够区分 KLF6/AhR 靶基因中的功能性 NC-XRE 与非功能性 NC-XRE。我将检查 KLF6/AhR 蛋白质-蛋白质相互作用，识别相互作用所需的蛋白质元件，并表征 KLF6/AhR 结合所需的关键 NC-XRE DNA 基序元件。人们普遍认为，接触二恶英后，AhR 是产生毒性所必需的。然而，人们对 AhR 驱动这种毒性发展的机制知之甚少。正如我们的数据表明，NC-XRE 代表了二恶英暴露后 AhR 的一种新型信号级联，这项工作将为二恶英毒性与 AhR 相关的机制提供新的见解，同时揭示用于识别和诊断的新型预后和诊断工具。治疗与 TCDD 暴露相关的有害健康影响。